Scientific Program - EKMF

Scientific Program
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T he Annual Meeting of the Brazilian Biophysics Society, one of the largest
and most traditional gathering of Biophysicists in Latin America, occurs
during the second semester of each year. Now in its 43rd edition, the 2018
meeting will be held again at the Mendes Plaza Hotel, in the city of Santos (SP),
from September 27th to 30th. A rich program of 16 symposia, 4 plenary talks, and
2 poster sessions covering a broad range of topics in Biophysics will guarantee
the appropriate environment for productive discussions about developments
regarding Biophysics and its interfaces with Biochemistry, Medicine, Pharmacy,
among other disciplines.
World leaders in several areas of Biophysics, from at least 12 different countries
have already confirmed participation in our meeting this year! We will host
researchers from Argentina, Belgium, Canada, Denmark, England, France,
Germany, Israel, Portugal, United States, Uruguay and, of course, from Brazil,
which will be represented by colleagues from 10 different states. This shows
our broad representation in both national and international scenarios.
We believe our meeting is the perfect opportunity for researchers interested in
Biophysics and related fields to present their latest results to a very distinguished
audience. Therefore, we invite you to participate and to submit an abstract for
the poster sessions.
Antonio Costa-Filho President of SBBf
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Organizing Committee ............................................7 Scientific Committee ..............................................7 Sponsors ................................................................7 Congress Venue......................................................8 Criação e diagramção: ...........................................8 Program .................................................................9 Plenary Lectures .................................................. 13
Gronenborn, A. M. ....................................................... 14 Benoît Roux ............................................................... 14 Aníbal E. Vercesi ......................................................... 14 J. A. Bouwstra ............................................................ 15
Pre-symposium .................................................... 16 PSY1 - Sirius and the new opportunities in biophysics ........................................................17
Carla Polo ................................................................... 17 Ana Carolina de Mattos Zeri ......................................... 17 Maria Cristina Nonato .................................................. 18
Symposium .......................................................... 19 SY1 - Cell and model membranes Biophysics .......20
Adriana Fontes ........................................................... 20 Koji Kinoshita .............................................................. 20 Nuno C. Santos ........................................................... 20 Luis A. Bagatolli .......................................................... 21
SY2 - Central Nervous System Electrophysiology .... 22 Leonardo Cirqueira ..................................................... 22 Melina Pires da Silva ................................................... 22 Christopher Kushmerick............................................... 23 Ricardo M. Leão .......................................................... 23
SY3 - Lipid-protein interactions in mimetic systems ..................................................24
Thierry Granjon ........................................................... 24 Ambrosio ................................................................... 24 M. Prieto .................................................................... 25 M. Bolean ................................................................... 25
SY4 - Nanostructures and Drug Delivery ..............26 Renata F. V. Lopez ....................................................... 26 Marcelo Morales ........................................................ 26 Omar Mertins .............................................................. 26 Lígia N. M. Ribeiro ....................................................... 27
SY5 - Protein Complexes ......................................28 Ana Paula Valente ....................................................... 28 Elio A. Cino ................................................................. 28 Rachel Nechushtai ....................................................... 28 Carla Mattos ............................................................... 29
SY6 - Modern Methods in Computational Biophysics ............................................................30
Kaline Coutinho ........................................................... 30 Gerald Weber .............................................................. 30 Rafaela S. Ferreira ...................................................... 30 Roberto A. Chica ......................................................... 31
SY7 - Antimicrobial peptides ................................32 Catarina Cataldi ........................................................... 32 Yeny Y. P. Valencia ....................................................... 32 João Ruggiero Neto ..................................................... 33
Katia Regina Perez ....................................................... 33 Maria Elena de Lima .................................................... 33
SY8 - Photobiology and its application in Health Sciences................................................35
Renato E. de Araujo ..................................................... 35 Yoshimura, T. M. ......................................................... 35 Tayana Tsubone .......................................................... 36 João Paulo Tardivo ...................................................... 36 Beate Röder ................................................................ 37
SY9 - Structural and Unstructural Biology ............38 Garegin A. Papoian ...................................................... 38 Vladimir N. Uversky ..................................................... 38 Yraima Cordeiro .......................................................... 38 Pinheiro, Glaucia M.S. ................................................. 39
SY10 - Biophysics of the heart .............................40 Danilo Roman Campos ................................................ 40 Rosana A. Bassani ...................................................... 40 Micaela Lopez Alarcón ................................................ 41 Emiliano Medei ............................................................ 41
SY11 - Methods in Biophysics ..............................42 Paul C. Whitford .......................................................... 42 Marin van Heel ............................................................ 42 Fabio Cesar Gozzo ...................................................... 42 Roberto K. Salinas ...................................................... 43
SY12 - Nanostructures .........................................44 Alan B. Dalton ............................................................. 44 Leandro Ramos Souza Barbosa ................................... 44 Frédéric Frézard ........................................................... 45 Daniele Ribeiro de Araujo ............................................. 45
SY13 - Molecular Modeling and Dynamics ...........46 Guilherme Menegon Arantes ........................................ 46 Werner Treptow ........................................................... 46 M. Pickholz ................................................................ 46 Sergio Pantano ............................................................ 47
SY14 - Photodynamic Therapy .............................48 Juliana Ferreira Strixino ................................................ 48 Fabio Parra Sellera....................................................... 48 Renata Aparecida Belotto ............................................. 48 Silvia Cristina Nunez .................................................... 49
SY15 - Biophysics of Proteins ..............................50 Carlos Henrique I. Ramos ............................................ 50 Chuck S. Farah ............................................................ 50 Jorge F. B. Pereira ....................................................... 50 Chehín, Rosana Nieves ................................................ 51
SY16 - Cellular Biophysics ...................................52 Celso Caruso Neves .................................................... 52 Fernando Abdulkader ................................................... 52 Felipe de Souza Leite .................................................. 53 Adriano M. Alencar ...................................................... 53
Posters ................................................................. 54 Biomaterials .........................................................55
Alex H. Miller ............................................................... 55 Aline Amorim Graf ....................................................... 55 Denys E. S. Santos ...................................................... 55 Elisângela Belleti ........................................................ 56
Iseli L. Nantes Cardoso ................................................ 56 Juliana S Yoneda ........................................................ 57 Lucas Rodrigues de Mello............................................ 57 Lucivaldo R. Menezes .................................................. 58 Mario de Oliveira Neto ................................................. 58 Maurício S. Baptista .................................................... 59 Rafael G. Carvalho....................................................... 59 Rhiannon W. Harries .................................................... 59 Vinicius Carrascosa ..................................................... 60
Biomedical Application .........................................61 Raissa L. Oblitas ........................................................ 61 André L.S. Santos ....................................................... 61 Angela D. B. de Brito .................................................. 62 Camila Ramos Silva .................................................... 62 Catarina Cataldi ........................................................... 63 Débora C. K. Codognato .............................................. 63 Cilli, E. M. ................................................................... 64 Cabral, F. V .................................................................. 64 Haroldo de Lima P. Cravo ............................................. 65 Juliana Guerra Pinto .................................................... 65 Letícia Corrêa Fontana ................................................. 66 L. M. Rebelo ............................................................... 66 Luciana Maria Cortez Marcolino .................................. 66 Natalia K. Gushiken ..................................................... 67 Nayara S. Alcântara Contessoto ................................... 67 Nicole M. A. Chaparro ................................................ 68 Sa. Arsalani................................................................. 68 Soudabeh. Arsalani ..................................................... 69 Taline S. Almeida ......................................................... 69 Taline S. Almeida ......................................................... 70 Juliana Ferreira Strixino ................................................ 70 Guilherme S. Ramos .................................................... 71
Biomembranes .....................................................72 Andresa Messias da Silva ............................................ 72 Bruna Renata Casadei ................................................. 72 Favarin, B. F. ............................................................... 73 C.R. SIMÕES ............................................................... 73 C. R. Ferreira ............................................................... 74 VESCHI E. A. ............................................................... 74 Fernando Freitas de Lima ............................................. 75 Frederico J. S. Pontes .................................................. 75 Gabriel S. Vignoli Muniz ............................................... 76 Sebinelli, H. G. ............................................................ 76 Helena Couto Junqueira ............................................... 76 Isabela Moreira-Silva ................................................... 77 Jefferson C Rodrigues ................................................ 77 Kenneth M. F. Miasaki .................................................. 78 Ludmilla D. Moura ....................................................... 78 L. H. Andrilli ................................................................ 79 Maressa Donato .......................................................... 79 Maria C. Oliveira ......................................................... 80 Natalia Alvarez ............................................................ 80 Natália B. Leite ............................................................ 81 Natália F. de Oliveira .................................................... 81 Strazza Junior PS ....................................................... 82 Priscilla Freddi ............................................................. 82 Raphael de A. N. Gomes ............................................. 83 Romildo A. Nogueira .................................................. 83
Tayana Mazin Tsubone ................................................. 83 Yan M. H. Gonçalves ................................................... 84 Yeny Y. P. Valencia ..................................................... 84 Virginia Sara Grancieri do Amaral ................................. 85
Biomembranes .....................................................86 Ana Paula R. Povinelli .................................................. 86 Anacleto Silva de Souza ............................................... 86 Anderson F. Sepulveda ................................................. 86 Balan, A. ..................................................................... 87 Aryane A. Vigato ......................................................... 87 Raphael Dias de Castro ............................................... 88 Amantino C. F ............................................................. 88 Lígia N. M. Ribeiro ....................................................... 89 Fabiana V. Diasa .......................................................... 89 Franciele Garcia Baveloni ............................................. 90 Gabriel Zazeri .............................................................. 90 Gianella Facchin1 ........................................................ 90 Giovana Firpo .............................................................. 91 Gonçalves G. E. G ...................................................... 91 Isabele Ap. S. de Campos ............................................ 92 João Hermínio Martins da Silva ................................... 92 Juliana Damasceno Oliveira ......................................... 93 Kelli Cristina Freitas Mariano ........................................ 94 Larissa D. da Silva....................................................... 94 Luciana Guimarães Munhoz ......................................... 95 Matheus P. Pinheiro ..................................................... 95 Mayk C. Ramos ........................................................... 96 Mayra C. G. Lotierzo ................................................... 96 Nascimento M. H. M .................................................... 96 Monique Lemos .......................................................... 97 Naially C. de Faria ........................................................ 97 Pablo V. M. Reis .......................................................... 98 Cordeiro Lima P. F ...................................................... 98 Bárbara Malheiros ....................................................... 99 Sandra B. N. Agostini .................................................. 99 Sandra M. G. Dias ..................................................... 100 Castro, Simone R. de................................................. 100 Viviane Corrêa Santos ............................................... 101 Amantino C. F ........................................................... 101
Molecular Mechanisms of Disease ..................... 102 Andreia Laura Prates Rodrigues ................................. 102 Artur S. Miranda ........................................................ 102 Marinonio L. Cornélio ................................................ 103 Osias B. S. Filho ........................................................ 104 Paula Rhana .............................................................. 104 Victor Barbosa .......................................................... 105 Diogo B. Peruchetti.................................................... 105 Ainhoa Rodriguez de Yurre ......................................... 106
Nucleic acids structure and functions ................. 107 Erik de Oliveira Martins .............................................. 107 Izabela F da Silva ....................................................... 107 Mateus Rodrigues Leal ............................................. 107 Pâmella Miranda ....................................................... 108 Vinícius Fernandes .................................................... 108 Vinícius G. Contessoto .............................................. 109 Vivianne Basílio Barbosa ............................................ 109
Protein Folding Misfolding and Unfolding ........... 111 Dayanne P. Rosa ........................................................ 111
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Fernando B. da Silva ................................................. 111 Gabriel G. Slade ........................................................ 112 Guilherme G. Costa ................................................... 112 Scanavachi, G. .......................................................... 113 Luciano Censoni ...................................................... 113 Mariana Chaves Micheletto ........................................ 113 Freire Ribeiro, M ........................................................ 114 Mariana P. Cali .......................................................... 114 Nelson A. Alves ......................................................... 115 Paulo Ricardo Mouro ................................................. 115 Rose M Carlos ......................................................... 115 Yulli M. F. Passos....................................................... 116
Protein Structure dynamics and Functions ......... 117 Agnes Magri ............................................................. 117 Anderson S. Pinheiro ................................................. 117 André Anversa Oliveira Reis ....................................... 118 Angie Dávalos ........................................................... 118 Antonio Victor B. Vasconcelos ................................... 118 Atílio Tomazini Júnior ................................................. 119 Penna, B. R. .............................................................. 119 Bianca Rizo ............................................................... 120 Bruno P. O. Santos ..................................................... 120 Camila A. Yamada ..................................................... 121 Camila F. T. Pontes ................................................... 121 Carmen Domene ....................................................... 121 Carolina T. A. Ferreira ................................................ 122 Catharina dos S. Silva................................................ 122 Claudemir Oliveira Souza ........................................... 123 Fabio C. L. Almeida ................................................... 123 Felipe A. Otsuka ........................................................ 124 Fernanda Paiva .......................................................... 124 Fernando T. Tanouye ................................................. 124 Fredderico C. Machado .............................................. 125 Gabriel Ernesto Jara .................................................. 125 Gabriel R. Sousa ....................................................... 125 Giovana C. Guimarães ............................................... 126 Glauce M. Barbosa .................................................... 126 Helder V. Ribeiro Filho ................................................ 127 Iara Aimê Cardoso ..................................................... 127 Ingrid B. S. Martins ................................................... 128 Isabella O. L. ............................................................. 128 Isis Sebastião............................................................ 129 Sarmento J. O .......................................................... 129 Jéssica A. Tedesco .................................................... 129 Jéssica M. de Sá ....................................................... 130 José Antonio Fiorote .................................................. 130 Juliana Raw .............................................................. 131 Juscemácia N. Araujo ................................................ 131 Karoline Sanches ...................................................... 131 Larissa G. Maciel ...................................................... 132 Bartkevihi, L. ............................................................. 132 Leonardo Boechi ....................................................... 133 Leonardo Cirqueira, .................................................. 133 Letícia Stock ............................................................. 134 Ramos, L. ................................................................. 134
Lilia Iriarte ................................................................. 135 Luan C. Marques ....................................................... 135 Lucas Bleicher .......................................................... 136 Lucas Carrijo de Oliveira ............................................ 136 Lucianna Helene Santos ............................................ 137 Luis Felipe Santos Mendes ........................................ 137 Luiz Fernando de C. Rodrigues ................................... 138 Bertozo L. C .............................................................. 138 Marcelo Querino Lima Afonso ................................... 139 Mariana A. Ajalla Aleixo ............................................ 139 Batista, M. R. B. ....................................................... 139 Marilia L. Cirqueira .................................................... 140 Marina G. Fontes ....................................................... 140 Marjorie C. L. C. Freire ............................................... 141 Miguel de S. Andrade ................................................ 141 Naiá Porã Santos ....................................................... 141 Natália A. Fontana ..................................................... 142 Neli Fonseca ............................................................. 142 Olívia Teixeira ............................................................ 143 Paola Silva ............................................................... 143 Paulo S. L. Beirão ..................................................... 143 Luccas, P. H. ............................................................. 144 Priscila dos S. Bury ................................................... 144 Rafael E. O. Rocha .................................................... 145 Rafael F. Soares......................................................... 145 Raissa Ferreira Gutierrez ............................................ 146 Ricardo A. P. Pádua ................................................... 146 Beserra S. dos S. Author .......................................... 147 Tábata Renée Doratioto ............................................. 147 Thainá Miranda ......................................................... 147 Thamires Q. Froes ..................................................... 148 Thirupathi R. Soudherpally ......................................... 148 Ximenes V. F ............................................................ 149 Vanessa S. Silva ........................................................ 149 Víctor U. Antunes ...................................................... 150 Victor Lopes Rangel ................................................. 150 Victoria Oakes ........................................................... 151 Vinícius M. Oliveira .................................................... 151 Vitor B. Machado ....................................................... 151 Pedro, R. P. ............................................................... 152 Marcus V. C. Cardoso ............................................... 153 Vitor B. P. Leite .......................................................... 153
Miscellaneous .................................................... 154 Felipe Curtolo ............................................................ 154 Marlon Augusto P. De Almeida .................................. 154 Nataly Herrera ........................................................... 154 Rafael C. Marchi ........................................................ 155 Rômulo P. Tenório ...................................................... 155 Amanda Schukarucha Gomes .................................... 156 Ana Carolina M. Figueira ............................................ 156 Telma Lisbôa Nascimento .......................................... 157 R. V. Maximiano ........................................................ 157 Fernando de Mesquita Júnior ..................................... 158
Alphabet Index .................................................. 159
Organizing Committee
Scientific Committee
• Antonio José da Costa - USP • Eneida de Paula - UNICAMP • Leandro R S Barbosa - USP
• Rosangela Itri - USP • Cynthia Bando - Secretária Geral
• Alessandro Nascimento - USP • Antonio José da Costa Filho - USP • Christopher Kushmerick - UFMG • Emiliano Medei - UFRJ • Eneida de Paula - UNICAMP • Ernesto Caffarena - FIOCRUZ
• Leandro R. S. Barbosa - USP • Lucas Bleicher - UFMG • Paulo Bisch - UFRJ • Pietro Ciancaglini - USP • Rosangela Itri - USP
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Congress Venue
Mendes Plaza Hotel Complexo Diamante Av. Marechal Floriano Peixoto 42 Gonzaga, Santos, SP Phone: + 55 13 3208-6400 www.mendesplaza.com.br
Criação e diagramção:
16:00 - 17:30
PSY1 - Pre-symposium: Sirius and the new opportunities in biophysics Chair: Chuck Farah (USP)
PSY1.1 – Carla Polo (LNLS): From macromolecules to cells: 3D imaging using coherent X-ray diffraction at Cateretê beamline (Sirius)
PSY1.2 – Ana Carolina de Mattos Zeri (LNLS): Macromolecular Crystallography at Sirius – frontiers and challenges for data collection and sample preparation at Manacá beamline
PSY1.3 – Maria Cristina Nonato (USP): X-ray crystallography fragment screening. Opportunities for Manacá beamline (Sirius)
16:30 - 18:00 Registration
18:00 - 19:00 PL01 – Opening lecture: Angela Gronenborn (Univ. Pittsburgh): “Not so crystal clear – crystallins and cataract” Chair: Antonio José Costa Filho (USP)
19:00 – 21:00 Poster session
SY1 – Cell and model membranes Biophysics Chair: Karin Riske (UNIFESP)
SY2 – Central Nervous System Electrophysiology Chair: Jader Cruz (UFMG)
SY1.1 – Adriana Fontes (UFPE): Optical Tweezers: Shining Light to Cell Biology
SY2.1B – Leonardo Cirqueira Pimentel (UNB): State-dependant affinity for general anesthetics dictates channel sensitivity
SY1.2 – Koji Kinoshita (Univ. South. Denmark): Red Blood Cell Membrane: Mechanical Property Change and Cytoskeletal Network Reformation Under Photosensitizer-mediated Oxidative Stress of Cis-Porphyrin
SY2.2 – Melina Pires (USP): Nitrergic Modulation of Magnocellular Neurons from the Supraoptic Nucleus
SY1.3 – Nuno Santos (Univ. Lisbon): Arterial hypertension and stroke patients present Higher erythrocyte adhesion forces, contributing to cardiovascular risk
SY2.3 – Christopher Kushmerick (UFMG): Ionotropic actions of Metabotropic Glutamate Receptors
SY1.4 – Luis Bagatolli (Univ. Nac.Córdoba): The cell as a gel: materials for a conceptual discussion
SY2.4 – Ricardo Leão (USP): Creating rest and unrest: distinct ways to create stable and unstable membrane potential in neurons
10:30 - 11:00 Coffee Break
Chair: Pietro Ciancaglini (USP)
SY4 – Nanostructures and Drug Delivery Chair: Daniele R. Araújo (UFABC)
SY3.1 – Thierry Granjon (Univ. Lyon): Cardiolipin and mitochondrial proteolipidic platforms: toward a mitochondrial raft-like organization?
SY4.1 – Renata Lopes (USP): Iontophoresis impact on topical delivery of nanoparticles
SY3.2 – André Luis Ambrosio (USP): Human MPC2 as an autonomous membrane transporter
SY4.2 – Marcelo Morales (UFRJ): Nanoparticle- mediated gene delivery for the lung
SY3.3 – Manuel Prieto (Univ. Lisbon): Surface crowding and membrane remodeling by IAPP
SY4.3 – Omar Mertins (UNIFESP): Development of functional Nanoparticles for delivery of bioactives
SY3.4 – Mayte Bolean (USP): Biophysical aspects of biomineralization
SY4.4 – Ligia Nunes de Morais Ribeiro (UNICAMP): Stability study of Drug Delivery Systems Based on Nanoparticle Tracking Analysis
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SY6 – Modern Methods in Computational Biophysics
Chair: Lucas Bleicher (UFMG)
SY5.1 – Ana Paula Valente (UFRJ): Conformational diversity in protein complexes: NMR studies of Allergens Bet v 1 and Fag s 1 with ligands and DIII of Dengue virus E protein with antibodies
SY6.1 – Kaline Coutinho (USP): Theoretical Studies of Electronic Structure of Molecules Interacting with Phospholipid Bilayers
SY5.2 – Elio Anthony Cino (UFMG): Role of Hydrogen Bonds in P53 Core Domain Stability and Aggregation
SY6.2 – Gerald Weber (UFMG): Mesoscopic models as a Tool for Probing molecular interactions in DNA and RNA
SY5.3 – Rachel Nechushtai (Hebrew Univ.): The role played by the labile 2Fe-2S clusters of neet proteins in controling their structure, function and folding
SY6.3 – Rafaela Ferreira (UFMG): Rational Drug Design Towards New Treatments For Neglected And Emerging Diseases
SY5.4 – Carla Mattos (Northeastern Univ.): Ras dimerization and allosteric modulation of GTP hydrolysis linked in the presence of Raf-RBD and the membrane
SY6.4 – Roberto Chica (Univ. Ottawa): Rational Design of Proteins that Exchange on Functional Timescales
17:30 - 18:30 PL02 -Plenary – Benoit Roux (Univ. Chicago): Molecular Dynamics Studies of P-type ATPase Ion Pumps Chair: Carla Mattos (Northeastern Univ)
18:30 - 19:30 SBBf General Assembly
SATURDAY Sep 29th
Chair: Silvia Nunez (Un. Brasil)
SY7.1A – Catarina Cataldi: Zika NS2B epitope as a candidate for diffential diagnosis – from Poster
SY8.1 – Renato Araújo (UFPE): Optical therapy and medical diagnosis assisted by metallic nanoparticles
SY7.1B – Yeny Yaneth Pillco Valencia: Computational simulation of popg aggregates in the presence of antimicrobial peptide LL-37 – from Poster
SY7.2 – João Ruggiero (UNESP): Lipid-packing perturbation induced by pH-responsive peptide in lipid membranes: effect of pH and peptide chemical modification.
SY8.2A – Tania M Yoshimura (IPEN): Biochemical changes in serum of obese mice related to photoac- tivation of brown adipose tissue
SY8.2B – Tayana Tsubone (USP): How does cell membrane respond to PDT?
SY7.3 – Katia Perez (UNIFESP): Using biomimetical membranes to study the mode of action of peptides
SY8.3 – Joao Paulo Tardivo (Faculdade de Medicina do ABC): Is Photodynamic Therapy Efficient to Treat Osteomyelitis in Diabetic Foot?
SY7.4 – Maria Elena Lima (UFMG): A synthetic peptide derived from a toxin of the spider Phoneutria nigriventer: a promising candidate to a therapeutic drug to treat erectile dysfunction
SY8.4 – Beate Roder (Univ. Humbolt): Photodynamic inactivation (PDI) of biofilm forming microorganisms
SY9.1 – Garegin Papoian (Univ. Maryland): Assembly and Dynamics of Histone Oligomers and Nucleosomes
SY10.1 – Danilo Campos (UNIFESP): Keep your heart stressed: impaired superoxide production and pathogenesis of Chagas disease
SY9.2 – Vladimir Uversky (Univ. South Florida): Unusual Biophysics and Strange Biology of Intrinsic Disorder
SY10.2 – Rosana Bassani (UNICAMP): Proarrhyhmic effects of Doxorubicin in the myocardium
SY9.3 – Yraima Cordeiro (UFRJ): Prion protein and alpha-synuclein aggregation in vitro: characterization of therapeutic compounds and cofactors for protein misfolding
SY10.3 – Micaela Lopez Alarcón (UFABC): The role of NLRP3/Casp1/IL-1β axis in renal ischemia/reperfusion-induced cardiac electrical disturbances
SY9.4 – Glaucia Pinheiro (UNICAMP): NMR structure of the J-domain of co-chaperone Sis1 from Saccharomyces cerevisiae
SY10.4 – Emiliano Medei (UFRJ): Cardiac electrical and mechanical study on a type 2 diabetes experimental model
13:00 - 15:00 Lunch
15:00 - 17:00
SY11.1 – Paul Whitford (Northeastern Univ.): Quantifying collective dynamics in the ribosome
SY12.1 – Alan Dalton (Univ. Sussex): Biological functionalization of synthetic nanoparticles using novel 2D peptide assemblies
SY11.2 – Marin van Heel (Imp. College): Dynamic Structures of Biological Nano-Machines by Single- Particle Cryo-EM
SY12.2 – Leandro RS Barbosa (USP): Cubosomes as delivery for many drugs
SY11.3 – Fabio Gozzo (UNICAMP): Advances in Mass Spectrometry as a Structural Biology Tool
SY12.3 – Frederic Frezard (UFMG): Targeting the host-cell for improved therapy of leishmaniasis
SY11.4 – Roberto Salinas (USP): Solution NMR spectroscopy studies of the Na+/Ca2+exchanger
SY12.4 – Daniele Araújo (UFABC): Thermosensitive hydrogels and organogels: from supramolecular structure to new drug-delivery systems
17:30 - 18:30 PL03 – Plenary – Anibal Vercesi (UNICAMP): The redox nature of the mitochondrial permeability transition Chair: Mauricio da S. Baptista (USP)
18:30 - 20:30 Poster Session
8:30 - 10:30
SY14 – Photodynamic Therapy Chair: Martha Ribeiro (IPEN)
SY13.1 – Guilherme Arantes (USP): Unfolding metalloproteins with force: From simple models to quantum mechanical calculations
SY14.1 – Juliana Ferreira Strixino (UNIVAP): Photodynamic therapy in cutaneous Leishmaniasis
SY13.2 – Werner Treptow (UnB): Binding of Small Ligands to Two State Membrane Proteins
SY14.2 – Fabio Parra Sellera (USP): Antimicrobial photodynamic therapy in Veterinary Medicine
SY13.3 – Monica Pickholz (Univ. B. Aires): Contributions of Simulations to the Understanding of Drug Delivery Systems
SY14.3 – Renata Belotto (Hospital Perola Byington): Can the photodynamic therapy be an alternative to treat high grade cervical intraepitheli- al neoplasia?
SY13.4 – Sergio Pantano (Pasteur): Towards a Complete Coarse-Grained Force Field for Biosimulations
SY14.4 – Silvia Nunez (Univ. Brasil): Antimicrobial Photodynamic Therapy
11:00 - 12:00 PL04 – Plenary – Joke Bouwstra (Leiden): Lipid organization in the skin barrier in health and disease Chair: Eneida de Paula (UNICAMP)
12:00 - 14:00 Lunch
SY16 – Cellular Biophysics Chair: Gilberto Weismuller (UFRJ)
SY15.1 – Carlos Ramos (UNICAMP): Chaperone- assisted protein aggregate reactivation
SY16.1 – Celso Caruso (UFRJ): albumin handling in renal proximal tubule cells: central role in the progression of renal disease
SY15.2 – Chuck Farah (USP): Structural and Functional Studies on the Xanthomonadaceae Type IV Secretion System – a Bacteria Killing Machine
SY16.2 – Fernando Abdulkader (USP): Ion channels in the amplifying pathway of insulin secretion
SY15.3 – Jorge Pereira (UNESP): Are Ionic Liquids Good Protein Stabilizers?
SY16.3 – Felipe Leite (USP): post-translational ar- ginylation and inter-sarcomere dynamics in skeletal muscle myofibrils
SY15.4 – Rosana Chehin (CONICET-UNT): Biophysical characterization of doxycycline potential neuroprotective mechanism
SY16.4 – Adriano Mesquita (USP): Dynamics of cardiomyocytes obtained by the method of traction force microscopy
16:00 - 16:30 Closing and awards ceremony
16:30 Refreshments
Plenary Lectures
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Gronenborn, A. M.
Xi, Z., Whitley, M.J., 1Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States
βγ-Crystallins are long-lived eye lens proteins that are crucial for lens transparency and refractive power. Each βγ- crystallin comprises two homologous domains, which are connected by a short linker. γ-Crystallins are monomeric, while β-crystallins crystallize as dimers and multimers. In the crystal, human βB2-crystallin is a domain-swapped dimer while the N-terminally truncated βB1-crystallin forms a face-en-face dimer. Combining and integrating data from multi-angle light scattering, nuclear magnetic resonance, and small-angle X-ray scattering of full-length and terminally truncated human βB2-crystallin in solution, we show that both these βB2-crystallin proteins are dimeric, possess C2 symmetry, and are more compact than domain-swapped dimers. Importantly, no inter-molecular paramagnetic relaxation enhancement effects compatible with domain swapping were detected. Our collective experimental results unambiguously demonstrate that, in solution, human βB2-crystallin is not domain swapped and exhibits a face-en-face dimer structure similar to the crystal structure of truncated βB1-crystallin
PL02 - Molecular Dynamics Studies of P-type ATPase Ion Pumps
Benoît Roux1
Huan Rui1, Avisek Das1, Francisco Bezanilla1, M. Holmgren2, Pablo Artigas3, Robert Nakamoto4. 1Department of Biochemistry and Molecular Biology, The University of Chicago, 929 E57th St, Chicago, IL 60637, USA. 2Molecular Neurophysiology Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA. 3Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center, Lubbock, United States 4Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, PO Box 800886, 480Ray C. Hunt Drive, Charlottesville, VA 22908, USA
P-class ATPases ion pumps constitute a superfamily of cation transport enzymes, present both in prokaryote and eukaryote, whose members mediate membrane flux of all common biologically relevant cations. P-class pumps use ATP to transport ions against their electrochemical potential (they are also called
E1-E2-type ATPase). The Na/K pump transports three Na+ out of the cell by two K+ into the cell at the expense of the hydrolysis of one molecule of ATP. The sarcoplasmic reticulum Ca2+-ATPase (SERCA) pumps two Ca2+ from the cytosol of muscle cells to the sarcoplasmic reticulum by exchanging two H+. From crystallography, we have a remarkable series of snapshots showing how these enzymes look at different states of their transport cycle. Using molecular dynamics simulation, the string method with swarms- of-trajectories, and free energy methods, we seek to understand the conformational dynamics involved as such pump transits through conformational states revealed by x-ray crystallography, the nature of the coupling between the binding of ATP, phosphorylation, and the movements of charged species across the core of the protein, the stepwise voltage-sensitive steps, and the origin of the ion binding specificity associated with different conformational states.1-8 A special attention is given to the protonation state of ionizable residues during the pumping cycle.9
References: 1.H. Yu, I. M. Ratheal, P. Artigas & B. Roux. Protonation of key acidic residues is critical for the K-selectivity of the Na/K pump. Nat. Struct. & Mol. Biol. 18, 1159-1163, (2011). PMC3190665. 2.J. P. Castillo, H. Rui, D. Basilio, A. Das, B. Roux, R. Latorre, F. Bezanilla & M. Holmgren. Mechanism of potassium ion uptake by the Na(+)/K(+)-ATPase. Nat. Comm. 6, 7622, (2015). 3.H. Rui, P. Artigas & B. Roux. The selectivity of the Na(+)/K(+)-pump is controlled by binding site protonation and self-correcting occlusion. eLife 5, (2016). 4.A. Das, H. Rui, R. Nakamoto & B. Roux. Conformational transitions and alternating- access mechanism in the sarcoplasmic reticulum calcium pump. J. Mol. Biol., (2017). 5.Y. Chen & B. Roux. Constant-pH Hybrid Nonequilibrium Molecular Dynamics-Monte Carlo Simulation Method. J. Chem. Theo. Comp. 11, 3919-3931, (2015).
PL03 - THE REDOX NATURE OF THE MITOCHONDRIAL PERMEABILITY TRANSITION
Aníbal E. Vercesi1
1Department of Clinical Pathology, State University of Campinas, São Paulo, SP - email: anibal@ unicamp.br
In addition to be the cell’s powerhouse, mitochondria also contain a cell death machinery that include highly regulated processes such as the membrane permeability transition (MPT) and reactive oxygen species (ROS) generation. High levels of matrix Ca2+ stimulate ROS production and opening of a nonspecific inner membrane pore, the permeability transition pore (PTP). These conditions disrupt energy-linked mitochondrial functions and may promote cell death
either by apoptosis or necrosis. A large body of evidence indicates that mitochondrial redox imbalance and MPT are responsible for the development and progression of a series of pathologies such as cancer, diabetes, dyslipidemias, inflammatory diseases, hypertension, ischemia/reperfusion injury, neurodegenerative diseases, muscular dystrophy, drug toxicity and aging. Here, we focus on the contributions of our laboratory to the understanding of the redox nature of MPT and the participation of PTP opening in mitochondrial dysfunction in hypercholesterolemic mice and statins toxicity.
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP # 17/17728- 8), Conselho Nacional Desenvolvimento Científico e Tecnológico, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior and Fundação de Apoio ao Ensino, à Pesquisa e à Extensão (UNICAMP/FAEPEX).
PL04 - Lipid organization in the skin barrier in health and disease
J. A. Bouwstra 1Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 RC, Leiden, The Netherlands
The skin barrier function is primarily located in the stratum corneum (SC) comprised of corneocytes (dead cells) and intercellular lipids. The main lipid classes in the stratum corneum are ceramides, free fatty acids, and cholesterol. These lipids form two crystalline lipid lamellar phases, one with a repeat distance of 13 nm (The long periodicity phase, LPP) and the other with a repeat distance of 6 nm (The short periodicity phase, SPP)(1). The most prevalent lateral packing is orthorhombic(2). The composition and organization of these lipids are crucial for a proper skin barrier function. Especially the LPP is considered to be crucial for the skin barrier function. In our group we are interested in the lipid composition and organization and how this is related to the skin barrier, especially to understand the impaired skin barrier function in inflammatory skin diseases. When summarizing the deviation in lipid composition in this tissue, we observed in atopic eczema (most prevalent skin disease in Europe) a reduced chain length of free fatty acids and of ceramides and an increase in the level of unsaturation(3). These changes in lipid composition resulted in an altered lipid lamellar organization and a lower lipid fraction of adopting an orthorhombic packing. Therefore the lipid properties substantially deviate from normal human SC. In order to understand
why these changes in lipid organization were observed, simultaneously studies were performed studies using lipid model systems(4). These studies revealed that we are able to form the LPP as well as the SPP in these lipid model systems and that the lipids mainly form a dense orthorhombic lateral packing. When changing the ceramide and/or fatty acid composition it was shown that long chain free fatty acids play an important role in the formation of the dense orthorhombic lateral packing and that a reduction in the free fatty acid chain length or an increase in the degree of unsaturation reduced the lipid fraction adopting an orthorhombic packing (5). These changes in the lipid composition increased also the permeability of these lipid membranes. These changes in lipid organization and permeability induced by changes in lipid composition explained very well the deviation in lipid organization and barrier reduction in atopic eczema skin. Finally using neutron, X-ray diffraction and infrared spectroscopy studies detailed analysis of the arrangement of the lipids in the SPP and LPP has been examined and it was observed that that the lipids in the LPP have a very exceptional arrangement with small lipid droplets in a fluid phase(6, 7). In conclusion the lipids in the stratum corneum have a very exceptional arrangement and play a prominent role in the reduced skin barrier in inflammatory skin diseases.
References: 1. Bouwstra J A, Gooris G S, van der Spek J A, et al. Structural investigations of human stratum corneum by small-angle X-ray scattering. J Invest Dermatol 1991: 97: 1005-1012. 2. Bouwstra J A, Gooris G S, Salomonsdevries M A, et al. Structure of Human Stratum-Corneum as a Function of Temperature and Hydration - a Wide-Angle X-Ray-Diffraction Study. International Journal of Pharmaceutics 1992: 84: 205- 216. 3.van Smeden J, Janssens M, Kaye E C, et al. The importance of free fatty acid chain length for the skin barrier function in atopic eczema patients. Exp Dermatol 2014: 23: 45-52. 4. van Smeden J, Janssens M, Gooris G S, et al. The important role of stratum corneum lipids for the cutaneous barrier function. Biochim Biophys Acta 2014: 1841: 295-313. 5.Mojumdar E H, Helder R W, Gooris G S, et al. Monounsaturated fatty acids reduce the barrier of stratum corneum lipid membranes by enhancing the formation of a hexagonal lateral packing. Langmuir 2014: 30: 6534-6543. 6.Paz Ramos A, Gooris G, Bouwstra J, et al. Evidence of hydrocarbon nanodrops in highly ordered stratum corneum model membranes. J Lipid Res 2018: 59: 137-143. 7.Mojumdar E H, Gooris G S, Groen D, et al. Stratum corneum lipid matrix: Location of acyl ceramide and cholesterol in the unit cell of the long periodicity phase. Biochim Biophys Acta 2016: 1858: 1926-1934
16 17
PSY1.1 - FROM MACROMOLECULES TO CELLS: 3D IMAGING USING COHERENT X-RAY
DIFFRACTION AT CATERETÊ BEAMLINE (SIRIUS)
Carla Polo1
1Laboratório Nacional de Luz Síncrotron (LNLS- CNPEM), Campinas-SP, Brazil e-mail: carla.polo@ lnls.br
The Cateretê beamline at Sirius, the new Brazilian synchrotron light source will be dedicated to coherent and time-resolved scattering experiments. It will provide unique capabilities with cutting edge research tools that are non-existent today in Brazil, like 3D imaging with nanometer resolution. In this talk an overview of the main characteristics and new scientific potentialities of coherent diffraction imaging (CXDI), X-ray photon correlation spectroscopy (XPCS) and ultra-small angle X-ray scattering (USAXS) for studying biomaterials will be provided as well as the status of the Cateretê project and the bio-experimental devices. We will present how CXDI studies allows to investigate structures in a wide range of length scales. This will be illustrated by the imaging characterization of morphological features in different lignocellulosic materials which shed light to biomolecular deposition/ deconstruction mechanisms that are important to plant biology and fuel production.
PSY1.2 - Macromolecular Crystallography at Sirius - frontiers and challenges for data
collection and sample preparation at Manacá beamline
Ana Carolina de Mattos Zeri
Sirius is the new brazilian Synchrotron light source, under construction in Campinas, São Paulo state. It will be a 4th generation source, with various applications in Biophysics. Advanced imaging techniques will be available alongside new in-operando techniques. The MANACÁ beamline will be installed in a high- beta straight section of the Sirius storage ring and its main goal is to bridge the gap between conventional crystallography, which uses large single crystals, and single-molecule X-ray diffraction performed in free electron lasers. The beamline will have two experimental stations, for the performance of
state-of-the-ar t macromolecular microcrystallography employing current methods, with a beam focused in the low tens of micrometers, and nanocrystallography, with a submicron focus beam and sub miliradian divergence. The beamline will operate with X-ray energies between 5 to 20 keV with large emphasis on membrane proteins and supramacromolecular protein complexes (megadalton size), such as receptors, proteasomes, translation complexes (protein and DNA/RNA components), bacterial cell division machinery and eukaryotic transcription complexes, and will enable a range of automated screening assays for molecular fragments and natural products. The applications cover areas such as pharmaceutics, agriculture and cosmetics, and will enable the use and development of new techniques for the investigation of biological materials at the atomic level. Accelerated methods for small molecule ligand screening are paramount for the development of new bio-catalysts and drug candidates, and the proposed methods of data collection and data analysis for single or multiple crystals in both experimental stations of the Manaca beamline will contribute to advances in Biotechnology, akin to what h been done in current third- generation sources. Atomic details of the active sites of the proteins under study and potential new important ligand binding pockets, and their interactions with several ligand candidates must be obtained in a fast manner, for ever smaller crystals. Frequently, proteins are obtained in small quantities, derived from natural sources, and yield microcrystals that must be screened under many different conditions. Methods of serial data collection, employing micro and sub-micron beams, with crystals in plates or micro-chips are a reality in microfocus beamlines in 3rd and 4th genera SR sources, and will be available at the Microfocus station of Manaca. New methods such as the acoustic drop levitation, to be implemented in the Nanofocus station, will also be important for the understanding of these enzymes. Current platforms for data collection, storage and distribution are being tested at the UVX to simulate the needs of the new beam lines. Software tools for data treatment are being developed and the team is contributing to the international MXCube data collection software consor tium. The open source collaborative platforms MXCube and ISPyB are employed and maintained by a group of Synchrotron Laboratories staff members, and is aimed at speeding up and making the data acquisition and treatment, up to structure solution, a more straightforward process for the users.
PSY1 - Sirius and the new opportunities in biophysics
(Sirius)
Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP, Ribeirão Preto, SP ([email protected])
Fragment screening has emerged as a modern approach to identify novel lead compounds for drug development. Moreover, fragment screening can identify secondary sites in proteins that may have a biological function. This approach entails screening libraries of very small molecules that follow the “rule of three” criteria. The smaller and less complex nature of fragments increases the probability of binding to a target protein, resulting in higher hit rates and an efficient search of diverse chemical space. Hits identified from fragment screening can be efficiently developed by growing, merging, or linking them to produce drug leads. Today, an array of biophysical methods has been developed to rapidly identify weakly binding fragments to a target protein: SPR, DSF, NMR, among many others. Each method has advantages
and disadvantages in terms of sample consumption, degree of automation, and assay complexity. However, because the affinities of the initial fragments are often low, detection methods are pushed to their limits, leading to a variety of ar tifacts, false positives, and false negatives that too often go unrecognized. Investigators in industry and academia have overcome these challenges by taking advantage X-ray crystallography techniques, which provide structural information that enables rapid and efficient assessment of hits. Also, recent technical developments enabled screening on a timescale comparable to other techniques allowing its routine use in drug discovery. In this present work, we will discuss the details (hardware/software) of High throughput X-ray Crystallography Fragment Screening (HTXCFS), illustrated with examples of our latest experiments at XCHEM facility at Diamond Light Source, and will bring to attention of future users of Sirius our effor ts/ambitions to build at the Manacá beamline a world leader facility for the application of HTXCFS.
This work was supported by Fundação de Apoio à Pesquisa do Estado de São Paulo - FAPESP (2016/22929-0), (2017/26559-5)
SY1.1 - OPTICAL TWEEZERS: SHINING LIGHT ON CELL BIOLOGY
Adriana Fontes1 1Department of Biophysics and Radiobiology, Federal University of Pernambuco, Recife, PE - ([email protected])
Advances in the field of biophotonics have led to the development of new optical tools able to be applied to study diverse biological events. The dual nature of light has allowed extracting important morphological, rheological, and chemical information related to the cellular and molecular world of biological systems. The optical tweezer, as a biophotonic tool, has been used not only to capture and move small particles (from micrometers to nanometers in size) but also to measure its mechanical properties. The optical tweezer is a sensitive tool capable of: (i) measuring molecular and cellular biophysical parameters, without mechanical contact; (ii) detecting small variations in biological properties, and (iii) analyzing cells and biomolecules one by one providing individual measurements, rather than just average values, which may help to identify signatures of many biological conditions. In this talk, we will present the basic principles related to the optical tweezer and illustrate its potential detailing biophotonic applications, such as those developed for the study of red blood cells and protozoa.
This work was supported by CNPq, CAPES, and FACEPE. It is also linked to the National Institute of Science in Photonics (INCT-INFo).
SY1.2 - Red Blood Cell Membrane: Mechanical Property Change and Cytoskeletal Network
Reformation Under Photosensitizer-mediated Oxidative Stress of Cis-Porphyrin
Koji Kinoshita
Department of Molecular Medicine, Faculity of Health Science, University of Southern Denmark, Odense, Denmark
Red blood cell (RBC) membrane has highly deformable and resilient property, due to a cytoskeletal network consisting of proteins such as band 4.1R, spectrin, and actin. However, the connection between the cytoskeleton network and the mechanical properties of RBC membranes – i.e., “signaling” – is still not well understood. This is because of lack of comprehensive study to understand the complex of the signaling. Recently, our research group found that a photosensitizer-mediated
oxidative stress of cis-porphyrin (CisDiMPyP) could induce a continuous change of RBC deformations, depending on irradiation degree, until finally causing hemolysis. To understand the signalling for the RBC morphological change under the condition, RBC shear modulus was measured by using micropipette manipulation techniques. The obtained results showed the increasing of RBC shear modulus from 6.7 x 10-6 to 11 x 10-6 N/m dynamically or statistically depend on the photosensitizer effect. We hypothesize that cytoskeletal network reformation with the oxidative stress could cause these behaviors. Therefore, the formation of cytoskeletal network of band 4.1R was also investigated by using the Stimulated Emission Depletion (STED) microscopy, and successfully visualized the band 4.1R positions of 169 ± 13 and 224 ± 10 nm distance (corresponded with a reference result of contour length of spectrin dimers between anchoring positions). We consider that this further characterization of the RBC membrane with the signaling will help to understand more complicated cytoskeletal network of immune, cancer or even neuron cells for therapeutic development in the future.
SY1.3 - Arterial hypertension and stroke patients present higher erythrocyte adhesion forces,
contributing to cardiovascular risk
Filomena A. Carvalho1, Ana Filipa Guedes1, Carlos Moreira2, Ana Catarina Fonseca1,2, José B. Nogueira2, José M. Ferro1,2 1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal ([email protected]) and 2Hospital Santa Maria, Centro Hospitalar Lisboa Norte, Lisbon, Portugal
The increase of erythrocyte aggregation by high fibrinogen levels may be an indicator of cardiovascular risk. γ’ fibrinogen variant has been considered as a possible player in enhancing aggregation. Here, we assessed, at the single-cell level, the influence of fibrinogen on erythrocyte aggregation in essential arterial hypertension (EAH) and stroke, as well as how it constitutes a cardiovascular risk factor for these patients. We also aimed at understanding how γ’ fibrinogen is altered on these patients. Differences on fibrinogen- erythrocyte interaction and cell-cell adhesion forces were evaluated by atomic force microscopy (AFM)- based force spectroscopy, with cells from EAH patients, stroke patients and healthy blood donors. Results were correlated with patients’ clinical profiles. AFM data show that the work and force necessary for erythrocyte- erythrocyte detachment is higher for patients than for healthy donors, with these parameters further increasing
in both groups when higher fibrinogen concentrations are present. Fibrinogen-erythrocyte (un)binding forces were also higher for patients, when compared with the control group, despite a lower binding frequency. The results can be associated with changes in blood flow, due to transient bridging of two erythrocytes by fibrinogen, representing an important cardiovascular risk factor. γ’ fibrinogen may contribute for the increased risk in both diseases, as we demonstrate that its levels are increased in these patients’ blood. Our results may be relevant for potential future drug interventions to reduce erythrocyte aggregation and enhance microcirculatory flow conditions in cardiovascular patients.
Guedes et al. (2016) Nature Nanotechnol. 11, 687; Guedes et al. (2017) Nanoscale 12, 14897.
SY1.4 - The cell as a gel: materials for a conceptual discussion
Luis A. Bagatolli
Instituto de Investigación Médica Mercedes y Martín Ferreyra (INIMEC-CONICET-Universidad Nacional de Córdoba), Friuli 2434, 5016- Córdoba, Argentina.
In recent studies we established that glycolytic oscillations in non-dividing yeasts are tightly coupled with the global state of intracellular water, being this phenomenon scale independent. Specifically, we demonstrated that an optimum dynamic state of the major component of the cell cytosol, water, modulated by optimal levels of ATP and an optimally organized actin network is crucial to the emergence of the oscillations (1-3), supporting the view of a highly coherent and ordered cellular interior with properties similar to a responsive hydrogel. These results, which are difficult to conceptualize using canonical cell models based in
mass action kinetics and dilute solution theory, can be mechanistically depicted using an alternative theory called the Association-Induction Hypothesis, proposed by G. N. Ling in the 60’s (4). Additionally, in the same oscillating cells we also measured temporal oscillations in thermodynamic variables such as temperature, heat flux and volume. Oscillations in these variables have the same frequency as oscillations in the activity of intracellular metabolites, suggesting strong coupling between them. These results can be interpreted in light of a recently proposed theoretical formalism by T. Heimburg(5) in which isentropic thermodynamic systems can display coupled oscillations in all extensive and intensive variables, reminiscent of adiabatic waves. This interpretation suggests that oscillations may be a consequence of the requirement of living cells for a constant low entropy state while simultaneously performing biochemical transformations, i.e., remaining metabolically active. This hypothesis, which is in line with the view of the cellular interior as a highly structured and near equilibrium system where energy inputs can be low and sustain regular oscillatory regimes, challenge the notion that biological processes are essentially dissipative.
References: 1) Thoke HS, et al. (2015) Tight coupling of metabolic oscillations and intracellular water dynamics in Saccharomyces cerevisiae. PLoS One 10(2):e0117308. 2) Bagatolli, L.A. and Stock, R. P. 2016. "The cell as a gel: material for a conceptual discussion" Physiological Mini Reviews 9(5): 38-49 3) Thoke HS, Thorsteinsson S, Stock RP, Bagatolli LA, & Olsen LF (2017) The dynamics of intracellular water constrains glycolytic oscillations in Saccharomyces cerevisiae. Sci Rep 7(1):16250 4) Ling GN (2001) Life at the cell and below cell level. The hidden history of a fundamental revolution in biology (Pacific press). 5) Heimburg T (2017) Linear nonequilibrium thermodynamics of reversible periodic processes and chemical oscillations. Phys Chem Chem Phys 19(26):17331-17341
Leonardo Cirqueira1
1Department of Cell Biology, Biology Institute, University of Brasília, DF ([email protected])
Since their discovery, general anesthetics allowed a big change in medicine, making possible some procedures that were unconceived without this class of drugs. Despite the wide pharmacological descriptions, major advances in anesthesiology, the general anesthesia mechanism remains unknown. Electrophysiology studies show that the main targets of general anesthetics(GA) are the brain ion channels. There are two main points of view that concern the mechanism, the indirect hypothesis, which states that change in membrane properties affects channels, and the direct- site hypothesis, which assumes that direct and specific interactions between anesthetics and ion channels modulate their behavior. Through direct-site point of view, modulation of a protein can be seen as differential interaction between the two states, which suggest that there must be differences in most probable positions and in the number of ligands that neighbor the protein. The molecular target used here is the mammalian voltage-gated potassium channel Kv1.2, which has well characterized both open and closed states atomic structures and is positively modulated by GA. Using the atomic models, there were performed extended (> 200ns) molecular dynamics simulations of both ion channel conformations embed in a membranous system with fixed sevoflurane, isoflurane or propofol concentrations. From each simulation, the spatial mean probabilities of occurrence were acquired. In positive modulators (sevoflurane and isoflurane) there were differences between the open and closed states. In addition, simulations with propofol, an insensitive ligand, will check if the different interaction hypothesis also explain the insensitive cases. These results offer a new and simpler way to see ligand-modulated protein activation, and helps to explain a single component of a more complex event: the general anesthesia.
This work was supported by CNPq, FAPDF and CAPES.
SY2.2 - Nitrergic Modulation of Magnocellular Neurons from the Supraoptic Nucleus
Melina Pires da Silva1
Davi José de Almeida Moraes1, André de Souza Mecawi2, José Antunes Rodrigues1 and Wamberto Antonio Varanda1
1Department of Physiology, School of Medicine of Ribeirão Preto – University of São Paulo, SP ([email protected]). 2 Federal Rural University of Rio de Janeiro, Department of Physiological Sciences, Seropedica, Brazil.
The control of the excitability in magnocellular neurosecretory cells (MNCs) of the supraoptic nucleus has been attributed mainly to synaptic inputs from circunventricular organs. However, nitric oxide (NO), a gaseous messenger produced in this nucleus during isotonic and short-term hypertonic conditions, is an example of a modulator that can act directly on MNCs to modulate their firing rate. NO inhibits the electrical excitability of MNCs, leading to a decrease in the release of vasopressin and oxytocin. Although the effects of NO on MNCs are well established, the mechanism by which this gas produces its effect is, so far, unknown. Because NO acts independently of synaptic inputs, we hypothesized that ion channels present in MNCs are the targets of NO. To investigate this hypothesis, we used the patch-clamp technique in vitro and in situ to measure currents carried by hyperpolarization- activated and nucleotide-gated cation (HCN) channels and establish their role in determining the electrical excitability of MNCs in rats. Our results show that blockade of HCN channels by ZD7288 decreases MNC firing rate with significant consequences on the release of OT and VP, measured by radioimmunoassay. NO induced a significant reduction in HCN currents by binding to cysteine residues and forming S-nitrosothiol complexes. These findings shed new light on the mechanisms that control the electrical excitability of MNCs via the nitrergic system and strengthen the importance of HCN channels in the control of hydroelectrolyte homeostasis.
This work was supported by CAPES, FAPESP and CNPq
SY2 - Central Nervous System Electrophysiology SY2.3 - Ionotropic actions of metabotropic glutamate receptors.
Christopher Kushmerick (1,2)
Everton dos Santos e Alhadas(1), Ana Maria Bernal Correia(1), Jennifer Diniz Soares Guimarães(1)
1) Programa de Pós-Graduação em Fisiologia e Farmacologia – ICB – UFMG. 2) Departamento de Fisiologia e Biofísica – ICB – UFMG
Metabotropic glutamate receptors are G-protein coupled receptors widely expressed in the central nervous system. Binding of ligand to these receptors activates signaling pathways leading to changes in neuronal function. Here, we describe electrophysiological actions of mGluR receptors that increase neuronal excitability resulting in functional changes to neurotransmission between the neurons involved.
SY2.4 - Creating rest and unrest: distinct ways to create stable and unstable membrane potential in
neurons.
Ricardo M. Leão 1Department of Physiology-FMRP, University of São Paulo, Ribeirão Preto, SP ([email protected]).
Central neurons usually have a stable resting membrane potential, firing action potentials only in response to synaptic stimulation. However, some neurons have unstable membrane potentials leading to the spontaneous firing of action potentials. But some neuronal types present both types of behavior, a silent or quiet mode, with a stable resting membrane potential, and an active mode, which fires action potentials spontaneously. Fusiform and cartwheel neurons from the dorsal cochlear nucleus present such behavior, with 50% of fusiform neurons present an active behavior and more than 80% of cartwheel neurons firing spontaneous action potentials at rest. Moreover, cartwheel neurons can present biostability, a situation where a neuron transits spontaneously between 2 states. Here I will present the ionic mechanisms that these neurons use to produce the quiet and active states, and show that specific ion channels have distinct effects on the creation of quiet and active neurons in a neuronal population.
This work was supported by FAPESP (2012/09421-6, 2016/01607-4)
SY3.1 - CARDIOLIPIN AND MITOCHONDRIAL PROTEOLIPIDIC PLATFORMS: TOWARD A
MITOCHONDRIAL RAFT-LIKE ORGANIZATION?
Department of Chemistry and Biochemistry, University of Lyon, ICBMS UMR 5246-CNRS, France
Aim/hypothesis - Cardiolipin (CL) is a mitochondrial phospholipid predominantly found in the inner mitochondrial membrane and known to be crucial in several metabolism processes and in apoptosis. The existence of functional CL platforms formed at the inner mitochondrial membrane under specific protein binding was often evocated and discussed. We hypothesise that the binding of two mitochondrial proteins, involved in the metabolism processes and known to be associated with CL, the mitochondrial isoforms of creatine kinase (mtCK) and of nucleoside diphosphate kinase (NDPKD) leads to such proteolipidic platforms.
Research design and Methods - Simplified biomimetic systems made of CL-containing liposomes or monolayers were prepared to test mtCK and NDPKD binding effect on membrane fluidity, lipid organisation and protein structure. Protein structural modifications upon membrane binding and effects on liposome membrane fluidity were analysed by combining both intrinsic or extrinsic fluorescence (using Laurdan as fluorescent probe) and infrared spectroscopy. CL organisation within monolayers after interaction with proteins was visualised with Brewster angle microscopy.
Results - Fluorescence and infrared spectroscopy approaches on liposomes indicated changes both in protein structure and phospholipid physical state upon the protein-lipid interaction. Laurdan fluorescence measurements indicated that protein binding to liposomes changed the phospholipid liquid-crystalline state to a more rigid state. Brewster angle microscopy investigations provide, for the first time, images indicating that both mtCK and NDPKD binding induced cluster formation on CL monolayers consolidating and changing the morphology of the interfacial film. Altogether, those findings support the hypothesis of the existence of proteolipidic platform in the inner mitochondrial membrane and suggest a major role of CL in the structuring of the mitochondrion.
This work was supported by University Lyon 1, CNRS and French Ministery of Research
SY3.2 - HUMAN MPC2 AS AN AUTONOMOUS MEMBRANE TRANSPORTER
Ambrosio1
R. Nagampalli1, J.E. Quesñay1, D. Adamoski1, Z. Islam1, J. Birch2, H. Sebinelli3, R. Girard4, C. Ascenção1, A. Fala1, B. Pauletti1, S. Consonni1, J. Oliveira1, A. Silva1, K. Franchini1, A. Paes Leme1, A. Silber4, P. Ciancaglini3, I. Moraes2, S. Dias1 & A. 1LNBio, CNPEM, Brazil(andre.ambrosio@lnbio. cnpem.br). 2MPL, DLS, England; 3FFCLRP, USP, Brazil; 4ICB, USP, Brazil
The active transport of glycolytic pyruvate across the inner mitochondrial membrane is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa heterotypic oligomer. Here, the recombinant production of human MPC through a co-expression strategy is first described; however, substantial complex formation was not observed, and predominantly individual subunits were purified. In contrast to MPC1, which co-purifies with a host chaperone, we demonstrated that MPC2 homo- oligomers promote efficient pyruvate transport into proteoliposomes. The derived functional requirements and kinetic features of MPC2 resemble those previously demonstrated for MPC in the literature. Distinctly, chemical inhibition of transport is observed only for a thiazolidinedione derivative. The autonomous transport role for MPC2 is validated in cells when the ectopic expression of human MPC2 in yeast lacking endogenous MPC stimulated growth and increased oxygen consumption. Multiple oligomeric species of MPC2 across mitochondrial isolates, purified protein and artificial lipid bilayers suggest functional high- order complexes. Significant changes in the secondary structure content of MPC2, as probed by synchrotron radiation circular dichroism, further supports the interaction between the protein and ligands. Our results provide the initial framework for the independent role of MPC2 in homeostasis and diseases related to dysregulated pyruvate metabolism.
Funded by grants and scholarships from FAPESP and The Wellcome Trust.
SY3.3 - Surface crowding and membrane remodeling by IAPP
M. Prieto1
J.C. Ricardo1, G. Scanavachi2, F. Fernandes1, R. Itri2 and A. Coutinho1,3
1CQFM-IN and IBB, Instituto Superior Técnico, Univ. Lisboa, Lisboa, Portugal (manuel.prieto@tecnico. ulisboa.pt), 2 Dept of Applied Physics, Univ. Sao Paulo, Sao Paulo, Brazil, and 3Dept of Chemistry and Biochemistry, Fac. Ciências, Univ. Lisboa, Lisboa, Portugal
Membrane-catalyzed amyloid fibril formation of human islet amyloid polypeptide (hIAPP) or amylin has been implicated as a mechanism by which hIAPP exerts its toxicity in type 2 diabetes. On the other hand, the membrane-bound aggregates of the rat variant of IAPP (rIAPP) are unable to progress into fibrillar structures since this is a non-amyloidogenic peptide. Here, we combined steady-state and time-resolved fluorescence methods and microscopy techniques to perform a comparative study on how anionic lipid membranes control the self-assembly of these two peptides. Our studies confirmed that membrane-catalyzed fibrillation of hIAPP results in fibril growth into the solution. However, the membrane-mediated self-assembly of rIAPP is confined to the lipid bilayer. To evaluate the impact of unlabeled rIAPP/lipid concentration on the membrane surface coverage reached by the peptide, FRET-based measurements of membrane binding of a tracer amount of fluorescently-labeled Atto488- rIAPP were combined with fluorescence anisotropy measurements of its membrane-bound oligomerization state. We found that progressive membrane saturation with rIAPP correlated with its ability in reducing the average size of anionic liposomes. Furthermore, a time- resolved emission spectra (TRES) study of Laurdan revealed that membrane binding and oligomerization of rIAPP produced an increased rigidity and surface dehydration in the vicinity of the probe that ultimately might be responsible for rIAPP ability in remodeling the lipid membranes.
This work was supported by the Bilateral Project FAPESP - FCT (FAPESP/20107/2014) and PPBI-POCI-01-0145- FEDER-022122. JCR acknowledges grant SFRH/ BD/95856/2013 from FCT. (Portugal)
SY3.4 - BIOPHYSICAL ASPECTS OF BIOMINERALIZATION
M. Bolean1
B.Z. Favarin1, H.G. Sebinelli1, E.A. Veschi1, M. Bottini2,4, M.F. Hoylaerts3. J.L. Millán4, P.Ciancaglini1. 1University of São Paulo, SP, Brazil (maytebolean@ usp.br) 2University of Rome Tor Vergata, Rome, Italy, 3University of Leuven, Belgium, 4Sanford Burnham Prebys Medical Discovery Institute, CA, USA.
During the process of endochondral bone formation, osteoblasts mineralize their extracellular matrix (ECM) by promoting the synthesis of hydroxyapatite (HA) seed crystals in the sheltered interior of membrane-limited matrix vesicles (MVs). Several lipids and proteins present in the membrane of the MVs mediate the interactions of MVs with the ECM and regulate the initial mineral deposition and posterior propagation. Among the proteins of MV membranes, ion transporters control the availability of phosphate and calcium needed for initial HA deposition. Phosphatases play a crucial role in controlling the inorganic pyrophosphate/inorganic phosphate ratio that allows MV-mediated initiation of mineralization. The lipid microenvironment can help in the nucleation process of first crystals and also plays a crucial physiological role in the function of MV-associated enzymes and transporters. The whole process is mediated and regulated by the action of several molecules and steps, which make the process complex and highly regulated. Liposomes and proteoliposomes, as models of biological membranes, facilitate the understanding of lipid–protein interactions with emphasis on the properties of physicochemical and biochemical processes. Here, we discuss the use of proteoliposomes as multiple protein carrier systems intended to mimic the various functions of MVs during the initiation and propagation of mineral growth in the course of biomineralization. We focus on studies applying biophysical tools to characterize the biomimetic models in order to gain an understanding of the importance of lipid– protein and lipid–lipid interfaces throughout the process.
Acknowledgements: FAPESP (2016/21236-0), CNPq (304021/2017-2,167497/2017-0) and CAPES.
SY4.1 - Iontophoresis impact on topical delivery of nanoparticles
Renata F. V. Lopez (USP)
The application of a constant electric current of low intensity, or iontophoresis, is traditionally used to administer drugs through the skin aiming their systemic action. With the advent of nanoparticles, the potential of iontophoresis to aid in the treatment of dermatological disorders, such as skin tumors and inflammation, has been explored in association with the nanoparticles. Nanoparticles can modify the movement of charged and non-charged hydrophilic molecules through the skin under the influence of iontophoresis enabling the targeting of drugs to specific regions of the skin. Iontophoresis may also facilitate the uptake of nanoparticles by tumor cells. Results will be presented regarding the influence of iontophoresis on the penetration of solid lipid nanoparticles, liposomes and immunoliposomes in skin and in skin tumors. Some surface properties of nanoparticles and their impact on iontophoresis and cellular internalization will be discussed. Finally, the effectiveness of treatment of skin tumors with nanoparticles by iontophoresis and by subcutaneous injection will be compared.
SY4.2 - Nanoparticle-mediated gene delivery for the lung
Marcelo Morales
Instituto de Biofísica Carlos Chagas Filho - UFRJ
Thymulin has been shown to present anti-inflammatory and anti-fibrotic properties in experimental lung diseases. We hypothesized that a biologically active thymulin analog gene, methionine serum thymus factor, delivered by highly compacted DNA nanoparticles composed of single molecule of plasmid DNA compacted with block copolymers of poly-L-lysine and polyethylene glycol (CK30PEG) that have been found safe in a human phase I clinical trial, may prevent lung inflammation and remodeling in a mouse model of allergic asthma. Thymulin plasmids were detected in the lungs of ovalbumin-challenged asthmatic mice up to 27 days after administration of DNA nanoparticles carrying thymulin plasmids. A single dose of DNA nanoparticles carrying thymulin plasmids prevented
lung inflammation, collagen deposition and smooth muscle hypertrophy in the lungs of a murine model of ovalbumin-challenged allergic asthma, leading to improved lung mechanics. In the present model of chronic allergic asthma, highly compacted DNA nanoparticles using thymulin analog gene modulated the inflammatory and remodeling processes improving lung mechanics.
SY4.3 - DEVELOPMENT OF FUNCTIONAL NANOPARTICLES FOR DELIVERY OF BIOACTIVES
Omar Mertins
Patrick D. Mathews, Ana C.M.F. Patta, Bianca B.M. Garcia.
Department of Biophysics, Federal University of Sao Paulo, São Paulo, SP. ([email protected])
Micro and nanoparticles are currently designed for plethora of biomedical, biological and environmental applications. The combination between polymers of different properties or between polymer and lipids has prompted the production of nano-delivery systems of specific physicochemical characteristics, which enable its structure stability during route of administration besides interaction to the expected target site, thus improving delivery and performance of transported bioactive compounds. In this sense, pH-responsive nanoparticles, composed by polysaccharides chitosan and alginate, where the two macromolecules respond differently to pH of application media thereby allowing both structural and surface charge modifications of the nanoparticles, actually improve delivery and pharmacological effect of transported drugs. Moreover, chitosan modified with arginine and associated to cationic liposomes has proven functional characteristics in interaction and complexation of plasmid-DNA, stability in cell culture media, besides promoting high transfection yields. Hence, this functional lipopolyplex boosts DNA delivery, which is under technologic scrutiny for genetic therapy. Indeed, the development of functional nanoparticles as drug and gene carriers is an emerging area, and further promising to medical applications.
This work was supported by the Sao Paulo Research Foundation (FAPESP: 2015/23948-5; 2016/13368-4).
SY4.4 - STABILITY STUDY OF DRUG DELIVERY SYSTEMS BASED ON NANOPARTICLE TRACKING
ANALYSIS
Verônica M. Couto1, Leonardo F. Fraceto2, and Eneida de Paula1
1Department of Biochemistry and Tissue Biology, University of Campinas, Campinas, SP ([email protected]) and 2Department of Environmental Engineering, São Paulo State University, Sorocaba, SP.
Evaluating the physicochemical stability of nanostructured colloids as drug delivery systems is a requirement to ensure their successfully application. However, assessing the structural properties of colloidal systems is difficult due to their intrinsic dynamism. Moreover, there is also a lack of in vitro techniques that can be directly correlated to the biological performance of colloids. Nanoparticle Tracking Analysis (NTA) has been exponentially applied in the in vitro colloidal
characterization. Based on its particle-by-particle approach, NTA provides size distribution and nanoparticle concentration data in real time. Lately, several works have demonstrated important relationship between nanoparticle concentration and biological performance of colloids. Given the importance of the nanoparticle concentration elucidation, we propose its use as mandatory analytical parameter in physicochemical stability of colloidal systems used in drug-delivery. The monitoring of nanoparticle concentration over time can also prematurely predict the degradation, erosion or disruption of the nanoparticles, which directly affect the colloid quality, and cannot be followed by changes in size distribution, as currently measured by Dynamic Ligth Scattering. Nanoparticle concentration is the parameter that bridges the abyss between the in vitro characterization and biological performance of colloidal systems, and therefore, should be monitored in the stability study of colloidal formulations.
This work was supported by the FAPESP (#14/25372-0 and #14/14457-5).
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SY5.1 - Conformational diversity in protein complexes: NMR studies of Allergens Bet v 1 and
Fag s 1 with ligands and DIII of Dengue virus E protein with antibodies
Ana Paula Valente1 1Department of Structural Biology, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil ([email protected])
We studied the structure and molecular dynamics of DIII, highlighting their role in the molecular mechanisms of Dengue virus infection and antibody recognition. We observed conformational exchange in the isolated DIII, in regions important for the packing of E protein dimers on the virus surface. Antibody binding not only removed the exchange regime in the epitope region, in a process reminiscent of conformational selection, but, somewhat surprisingly, also caused exchange in other parts of DIII through allosteric effects. We also studied the modulation of dynamics in allergens from Bet v 1 family observed upon ligand binding and the importance for antibody-epitope complex formation. Our data suggest that the ligand-binding cavity of the allergen and the related changes in structural dynamics, as a key structural element for allergenicity.
This work was supported by Conselho Nac. Des. Cient. Tecnológico (CNPq # 303785/2014-4; 426265) and by the Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro (E-26/202.902/2015; E-26/201.314/2016).
SY5.2 - Role of Hydrogen Bonds in P53 Core Domain Stability and Aggregation
Elio A. Cino
Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, MG ([email protected])
The functionality of tumor suppressor p53 is altered in over 50% of human cancers, and many patients exhibit amyloid-like buildups of aggregated p53. Understanding the initial steps that trigger pathogenic amyloid conversion of p53 is required for further development of cancer therapies. Despite having ~60% sequence identity, out results from Thioflavin-T kinetics experiments showed that the p53 core domain (p53C) aggregated faster, and to a greater extent than that of family members p63C and p73C, and was more
susceptible to pressure denaturation. The aggregation tendencies of p53C, p63C, and p73C were strongly correlated with their thermal and pressure denaturation midpoints. Molecular Dynamics simulations indicated specific regions of structural heterogeneity unique to p53C, which may be promoted by elevated incidence of exposed backbone hydrogen bonds (BHBs), reduced BHB strength, and increased water interaction with BHBs compared to p63C and p73C. Based on the higher sensitivity of p53C to pressure denaturation, and elevated number of vulnerable BHBs, we propose that water infiltration into the core of the p53C structure is a root cause of its instability. Using 3D sequence alignments, we have identified regions of structural vulnerability in p53C, suggesting new targetable sites for modulating p53C stability and aggregation, which is a potential approach to cancer therapy.
SY5.3 - THE ROLE PLAYED BY THE LABILE 2Fe-2S CLUSTERS OF NEET PROTEINS IN
CONTROLING THEIR STRUCTURE, FUNCTION AND FOLDING
Rachel Nechushtai1
Jose’ N. Onuchic2, Patricia A. Jennings3, Paolo Carloni4 and Ron Mittler5
1Alexander Silberman Life Science Institute, The Hebrew University, Jerusalem (rachel@maul. huji.ac.il) and 2Center for Theoretical Biological

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