Thirteenth Regional Symposium October 9-10 (Fri, Sat), 2020 Scientific Program and Abstracts

Higuchi Biosciences Center University of Kansas Lawrence, KS

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Welcome to GRASP 2020, the 13th Great Plains Regional Annual Symposium on Protein and Biomolecular NMR. This year has been unusual in many regards, but thanks to technology, we can meet and mark our thirteenth year of the meeting. This meeting has resulted from the enthusiastic participation and strong support from you, the biomolecular NMR community in the central Midwest. As has been the case in previous years, we anticipate that you will enjoy excellent presentations on research at the forefront of our field and will engage in interesting discussions about one another’s research. One of our main objectives for this meeting has been to create a forum that will stimulate ongoing interactions among colleagues and friends, as collaboration is of increasing importance for the advancement of science. We are glad that you are able to participate. It is also our hope that the opportunity to come together for further discussions will continue in the years to come. Thank you for helping to make GRASP NMR a success each year.

We are extremely grateful for the generous organizational support offered by the KU Higuchi Biosciences Center. They contributed greatly to the success of our GRASP meetings. Many thanks to them.

Thank you all for your participation. We hope you enjoy the meeting!

Best regards,

GRASP NMR 2020 Organizing Committee Moriah Beck, Associate Professor of Chemistry, Wichita State University Gabriel A Cook, Assistant Professor of Chemistry, Oklahoma State University Roberto De Guzman, Professor of Molecular Biosciences, University of Kansas Vincenzo Venditti, Assistant Professor of Chemistry, Iowa State University

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Thirteenth GRASP 2020 Great Plains Regional Annual Symposium on

Protein and Biomolecular NMR

University of Kansas, Lawrence, KS October 9−10, 2020

FRIDAY, OCTOBER 9, 2020 All lectures will be held via Zoom. Session 1 Chair: Dr. Moriah Beck 2:00-2:30 “Structural studies of the HIV-1 PBS-segment RNA: insights into virus:host

interactions in promoting viral infectivity” Xiao Heng, Associate Professor, Department of Biochemistry, University of Missouri 2:30-2:50 “Investigating the conformational dynamics in human RNA demethylase FTO” Balabhadra Khatiwada, Graduate Student, Department of Chemistry, Iowa State University 2:50-3:10 “Probing the Maturation of heterodimeric HIV-1 Reverse Transcriptase from its

homodimeric precursor by Double Electron‐Electron Resonance EPR Spectroscopy” Thomas Schmidt, Post-Doctoral Fellow, Laboratory of Chemical Physics, National Institutes of Health

3:10 – 3:20 Break Session 2 Chair: Dr. Vincenzo Venditti 3:20–3:50 “NMR-based structural biology approaches to understand molecular recognition and

conformational dynamics within large protein-nucleic acid systems"” Nicholas Reiter, Assistant Professor, Department of Chemistry, Marquette

University 3:50–4:10 “Dependence of Proline Isomerization on the Kinetics of Folding of Anthrax Lethal

Factor"” Wyel Halimeh, Undergraduate Student, Department of Chemistry Wichita State University

4:10-4:30 “RNA structure in reverse transcription initiation is important to maintain viral RNA

integrity” Thomas Gremminger, Graduate Student, Department of Biochemistry, University

of Missouri

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SATURDAY, OCTOBER 10, 2020

All lectures will be held via Zoom.

Session 3 Chair: Dr. Gabriel Cook 10:00 – 10:30 “Structural basis for Larp7 recognition of the noncoding 7SK RNA: an NMR-guided

approach” Catherine D. Eichhorn, Assistant Professor, Department of Chemistry, University of

Nebraska-Lincoln

10:30– 10:50 “Identification and Characterization of the Binding of Novel Small Molecules to Salmonella Tip Protein SipD”

Amritangshu Chakravarty, Graduate Student, Department of Molecular Biosciences, University of Kansas

10:50– 11:10 “Understanding Interactions of a Peptide Drug with Bile Salts under Simulated Gastrointestinal Conditions via NMR Spectroscopy”

Tahnee J. Dening, Postdoctoral Researcher, Department of Pharmaceutical Chemistry, University of Kansas

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13th Great Plains Regional Annual Symposium on Protein and

Biomolecular NMR

Abstracts of

Oral Presentations

October 9-10, 2020

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Xiao Heng , Ph.D. ~ Invited Speaker Associate Professor Department of Biochemistry University of Missouri Columbia, MO

Abstract

“Structural studies of the HIV-1 PBS-segment RNA: insights into virus:host interactions in promoting viral infectivity” Xiao Heng1*, Zhenwei Song1, Gatikrushna Singh2, and Kathleen Boris-Lawrie2. 1 Department of Biochemistry, University of Missouri, Columbia, 65211 2 Department of Veterinary and Biomedical Sciences, University of Minnesota, Saint Paul, MN, 55108 HIV-1 reverse transcription initiates at the primer binding site (PBS) in the viral genomic RNA (gRNA). Although the structure of the PBS-segment undergoes substantial rearrangement upon tRNALys3 annealing, the proper folding of the PBS-segment during gRNA packaging is important as it ensures loading of beneficial host factors, including human DHX9/RHA. Site-specific deuteration labeling NMR strategies were employed to study the structure of the PBS-segment. In combination with SAXS, we determined the solution structure of the PBS-segment RNA, which adopts a previously undefined three-way junction structure containing the primer activation stem (PAS), tRNA-like element (TLE) and tRNA annealing arm. Disruption of the PBS-segment three-way junction structure led to diminished reverse transcription products and thus reduced viral infectivity. Existence of the tRNA annealing arm prevents coaxial stacking of TLE on the PAS stem, and the bent helical structures of TLE and PAS are preferentially bound by RHA double-stranded RNA binding domain 1 (dsRBD1), suggesting a shape-dependent RNA recognition of dsRBD1. We also reported that DHX9/RNA helicase A (RHA) is recruited to the PBS-segment of the viral genomic RNA to enhance the processivity of reverse transcriptase. Overall, our structural

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studies of the PBS-segment RNA and its interaction with human RHA provide structural insights into the assembly of ribonucleoprotein complex for efficient reverse transcription.

Biography

Dr. Xiao Heng received her B.S degree in Nanjing University, China. She completed her Ph.D. and postdoc training with Dr. Michael F. Summers at HHMI/University of Maryland, Baltimore County. She joined the University of Missouri as an Assistant Professor in 2013, and was promoted to Associate Professor in 2020. She received NIH CRNA Career Transition Award in 2013 and MU Dr. Richard and Wallace Faculty Incentive Grant Award in 2014.

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Balabhadra Khatiwada

Graduate Student Department of Chemistry Iowa State University Ames, IA

Abstract

“Investigating the conformational dynamics in human RNA demethylase FTO” Balabhadra Khatiwada *1, Jeffrey A. Purslow1, Trang T. Nguyen1, and Vincenzo Venditti1,2* 1Department of Chemistry and 2Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA The fat mass and obesity-associated (FTO) protein is a non-heme Fe(II) and α-ketoglutarate-dependent AlkB dioxygenase that catalyzes oxidative demethylation of N6-methyladenosine (m6A) in single-stranded RNAs. The overexpression of FTO is associated with various human diseases such as obesity and cancer. Unlike other AlkB homologue proteins, FTO is comprised of two distinct domains; a catalytic N-terminal domain (NTD) and a structurally independent C-terminal domain (CTD) which has a unique fold and is found exclusively in FTO. AlkB proteins are known to be inherently dynamic and the conformational dynamics plays crucial role in the catalysis and substrate selectivity. In this work, we study the conformational dynamics and its role in the structure-function relationship of FTO using multidisciplinary approaches including enzyme catalysis, accelerated molecular dynamics (aMD) simulation as well as various NMR experiments; such as residual dipolar couplings (RDC), 15N and 13C-methyl relaxation based NMR experiments. Our data suggests that FTO is a highly dynamic protein on multiple timescales. In particular, majority of the loops on NTD exhibit ps-ns dynamics whereas the regions surrounding the active site and the NTD/CTD interface are dynamic on µs-ms timescale. Analysis of RDC data in conjunction with full-atom trajectories from aMD simulations shows that the protein assumes a high degree of conformational heterogeneity in the solution, which is likely to be important for substrate selectivity.

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Thomas Schmidt, Ph.D. Post-Doctoral Fellow Laboratory of Chemical Physics National Institutes of Health MD

Abstract

“Probing the Maturation of heterodimeric HIV-1 Reverse Transcriptase from its homodimeric precursor by Double Electron‐Electron Resonance EPR Spectroscopy” Thomas Schmidt*, John M. Louis and G. Marius Clore Laboratory of Chemical Physics, National Institutes of Health, MD

HIV type I (HIV-1) reverse transcriptase (RT) catalyzes the conversion of viral RNA into DNA, initiating the chain of events leading to integration of proviral DNA into the host genome. RT is expressed as a single polypeptide chain within the Gag-Pol polyprotein, and either prior to or following excision by HIV-1 protease forms a 66 kDa chain (p66) homodimer precursor. Further proteolytic attack by HIV-1 protease cleaves the ribonuclease H (RNase H) domain of a single subunit to yield the mature p66/p51 heterodimer. Here, we probe the spatial domain organization within the p66 homodimer using pulsed Q-band double electron-electron resonance (DEER) EPR spectroscopy to measure a large number of intra- and intersubunit distances between spin labels attached to surface-engineered cysteines. The relative orientation of the domains was modeled by simulated annealing driven by the DEER-derived distances resulting in a p66/p66′ asymmetric reverse transcriptase homodimer with the RNase H domain that is cleaved to generate p51 in the mature p66/p51 heterodimer is present in 2 major conformers. Subsequently, one conformer is fully solvent accessible thereby accounting for the observation that only a single subunit of the p66 homodimer precursor is susceptible to HIV-1 protease; hence a flexible, exposed linker between the RNaseH and connection domains in the open state of the p66′ subunit binds to the active site of protease in a configuration that is similar to that of extended

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peptide substrates. The derived structural model explains the maturation steps towards active HIV type I reverse transcriptase.

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Nicholas Reiter, Ph.D. ~ Invited Speaker Assistant Professor Department of Chemistry Marquette University Milwaukee, WI

Abstract

“NMR-based structural biology approaches to understand molecular recognition and conformational dynamics within large protein-nucleic acid systems” Nick Reiter*, Danyun Zeng, John Aguirre, and Dulmi Senanayaka Department of Chemistry, Marquette University, Milwaukee, WI Lysine-specific histone demethylase 1 (LSD1) is a conserved chromatin-remodeling enzyme that catalyzes the removal of mono- and dimethyl functional groups from Histone 3 proteins at lysine positions 4 and 9 (H3K4/K9). The N-terminus of human LSD1 consists of a long 170 amino acid tail with no predicted structural elements and the SWIRM domain, which forms a stable, compact tertiary fold situated atop the catalytic amine oxidase domain (AOD). Both the N-terminal tail of LSD1 and the SWIRM domain facilitate intermolecular protein interactions. Quantitative surface plasmon resonance demonstrate that LSD1-coREST’s nucleosome binding affinity is significantly improved upon the addition of the intrinsically disordered N-terminal tail, where the full length LSD1 in complex with CoREST binds unmodified nucleosomes containing a 187mer DNA sequence in a 1:1 stoichiometric manner at nanomolar affinity. NMR reveals the existence of an α-helical element that resides within a conserved region of LSD1’s N-terminus. The propensity for an α-helical conformational ensemble within the N-terminal region of LSD1 data is supported through combined chemical shift (CS)-Rosetta and explicit solvent molecular dynamics simulation. Further, X-ray crystallography corroborates the formation of a stable N-terminal α-helix of LSD1 and suggests a regulatory binding location of an α-helix-AOD interaction adjacent to the FAD cofactor site. The identification of structure, nucleosome binding, and intramolecular

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contacts within this LSD1-nucleosome system will be discussed. In addition, we will introduce an NMR-based approach to probe the dynamics of tRNA recognition by the large RNA-based RNase P holoenzyme. Biography

Dr. Nicholas (Nick) Reiter received his B.A. in chemistry from Carleton College, worked briefly at the Mayo Clinic (MN), and received his Ph.D. in Biochemistry in 2006 from University of Wisconsin-Madison with Prof. Sam Butcher. He was an NRSA NIH postdoctoral fellow with Prof. Alfonso Mondragon at Northwestern and subsequently joined the Biochemistry faculty at Vanderbilt University's School of Medicine in 2012, where he began his NIH funded research program. He joined Marquette University's Chemistry department in the fall of 2017 and his primary research interests are in the area of RNA-protein structure and function.

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Wyel Halimeh Undergraduate Student Department of Chemistry Wichita State University Wichita, KS

Abstract

“Dependence of Proline Isomerization on the Kinetics of Folding of Anthrax Lethal Factor”

Wyel Halimeh* and James G. Bann

Department of Chemistry, Wichita State University, Wichita, KS

The death that can come within a few days after exposure to the anthrax toxin from Bacillus anthracis is due to the presence and effects of anthrax lethal factor (LF). LF is a zinc metalloproteinase whose function depends upon the translocation of LF from the endosome of a host cell into the cytosol, where it cleaves mitogen-activated protein kinase kinases and disrupting cell signaling pathways. The translocation requires LF to unfold as it transits through the narrow channel of the pore, formed by the anthrax protective antigen (PA). Unfolding of LF has been shown to be pH-dependent, but little is known regarding the kinetics of unfolding and refolding of LF. Specifically, refolding of LF must occur fairly rapidly within the cell cytosol to prevent degradation of the protein by cellular proteases. The N-terminal PA binding domain of LF, LFN, has a single cis-proline residue (Pro166), and we hypothesized that this cis proline must isomerize to trans during the unfolding and translocation process and that refolding would be slow, and perhaps dependent on cellular prolyl isomerases for refolding. To this end, we have performed a detailed kinetic refolding/unfolding study of LFN from urea solutions. Our preliminary experiments indicate that LFN refolding occurs rapidly (within 1 second), suggesting that Pro166 does not isomerize to an appreciable extent in the unfolded state, or if it does isomerize, that isomerization back to cis is a fast process. The implications of these experiments on the mechanism of anthrax toxin lethality will be discussed.

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Thomas Gremminger Graduate Student Department of Biochemistry University of Missouri Columbia, MO

Abstract

“RNA structure in reverse transcription initiation is important to maintain viral RNA integrity” Thomas Gremminger*, Zhenwei Song, Juan Ji, Avery Foster, and Xiao Heng

Department of Biochemistry, University of Missouri, Columbia, 65211

Reverse transcription of HIV-1 initiates upon annealing of human tRNALys3 onto the primer binding site (PBS) in viral RNA (vRNA). Additional intermolecular interactions between tRNALys3 and vRNA have been reported, but their function in viral replication remains unclear. Here we show that abolishing one potential interaction, the A-rich loop: tRNALys3 anticodon interaction, led to a decrease in infectivity and reduced synthesis of reverse transcription products. The disruption of the A-rich loop: tRNALys3 anticodon interaction in the tRNALys3: vRNA complex structure was confirmed by nuclear magnetic resonance (NMR) experiments. In vitro primer extension experiments revealed that disruption of the A-rich loop: tRNALys3 anticodon interaction resulted in increased affinity for RT and enhanced reverse transcription efficiency. In the absence of dNTP, RNaseH activity of reverse transcriptase (RT) was observed, and the complex with the A-rich loop: tRNALys3 anticodon interaction underwent an accelerated degradation by RT. Consistently, loss of vRNA integrity was observed in virions containing A-rich loop mutations. These results suggest that the additional intermolecular interaction between tRNALys3 and vRNA protects the vRNA in the mature virions prior to entering a new host cell.

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Catherine Eichhorn, Ph.D. ~ Invited Speaker Assistant Professor Department of Chemistry University of Nebraska-Lincoln Lincoln, NE

Abstract

“Structural basis for Larp7 recognition of the noncoding 7SK RNA: an NMR-guided approach” Catherine D. Eichhorn* Department of Chemistry, University of Nebraska Lincoln. The 7SK ribonucleoprotein (RNP) is a major regulator of transcription during homeostasis and cell stress. Despite its critical function and biomedical significance, there are few structural insights into 7SK RNP function, in stark contrast to other regulatory RNPs like the spliceosome. 7SK RNA is constitutively bound to the La-related protein 7 (Larp7), an essential component of the 7SK RNP complex. Using solution NMR spectroscopy and binding assays, we identify the specific binding site between 7SK RNA and the Larp7 C-terminal domain and characterize RNA and protein dynamics using 13C spin relaxation and heteronuclear NOE experiments. Using solution NMR as a guide for construct design, we determine the 2.2 Å X-ray crystal structure of the Larp7 C-terminal domain in complex with 7SK RNA. We find that the Larp7 C-terminal domain contains an atypical RNA recognition motif that binds the 7SK RNA 3’ terminal stem-loop with high affinity and specificity. Combining structural and dynamics studies with molecular modeling, we propose a structural model for Larp7 binding to the 7SK 3′ end and a mechanism for 7SK RNP assembly. This work provides atomic-level insights that explain why deletion of the Larp7 C-terminus is associated with loss of specific binding to 7SK RNA and diseases such as gastric cancer.

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Biography

Catherine Eichhorn received her B.A. degree in Chemistry from Bryn Mawr College in 2007 and Ph.D. in Chemical Biology from the University of Michigan - Ann Arbor in 2012, where she worked with Prof. Hashim Al-Hashimi. She was an American Cancer Society Postdoctoral Fellow with Prof. Juli Feigon at the University of California - Los Angeles. She joined the University of Nebraska - Lincoln as an Assistant Professor in 2019.

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Amritangshu Chakravarty Ph.D. Candidate Department of Molecular Biosciences University of Kansas Lawrence, KS

Abstract

“Identification and Characterization of the Binding of Novel Small Molecules to Salmonella Tip Protein SipD”

Amritangshu Chakravarty2*, Nick J Lowe2, David Johnson1 and Roberto N. De Guzman2 1Computational Chemical Biology Core Facility, University of Kansas, Lawrence, KS 66047, USA 2Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045, USA

Pathogenic Gram-negative bacteria such as Salmonella use the Type III Secretion System (T3SS) to inject effector proteins into host eukaryotic cells. There has been an increasing emergence of multidrug resistant strains among the Gram-negative bacteria, and they all use the T3SS for infection. The T3SS is an attractive target for developing new classes of antimicrobials. The structural component of the T3SS is the needle apparatus, which consists of a base, an extracellular needle, a tip, and a translocon. The first step in developing new antimicrobials against the T3SS is the identification of novel small molecules that can bind to different protein components of the T3SS. Previously, some small molecules binding to the Salmonella tip protein SipD, were identified. Here we have screened small molecule and compound libraries using surface plasmon resonance (SPR), virtual screening and Rosetta. We identified 6 compounds that bind specifically to SipD. We have also used NMR to determine the residues and surfaces of SipD interacting with these compounds. This knowledge can be used in the future to design novel drug molecules that can combat multidrug resistance associated with the Gram-negative bacteria.

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Tahnee J. Dening, Ph.D. Postdoctoral Researcher Department of Pharmaceutical Chemistry University of Kansas Lawrence, KS

Abstract

“Understanding Interactions of a Peptide Drug with Bile Salts under Simulated Gastrointestinal Conditions via NMR Spectroscopy” Tahnee J. Dening1*, Justin T. Douglas2 and Michael J. Hageman1

1Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 2Nuclear Magnetic Resonance Core Laboratory, University of Kansas, Lawrence, KS Unfavorable properties of peptide drugs such as their susceptibility to gastrointestinal enzymatic degradation and low membrane permeability necessitate that these difficult-to-deliver molecules be administered via injection, which is undesirable from a patient perspective. Accordingly, there is major interest in developing peptide drugs for oral administration, however, the impact of the aqueous gastrointestinal environment on peptide drug behavior in solution and oral absorption (beyond enzymatic degradation) has been largely neglected; the interaction of peptide drugs with bile salts and bile salt mixed micelles under simulated gastrointestinal conditions has not been reported in the scientific literature. An improved understanding of peptide drug-bile salt interactions in solution is essential, especially given large inter- and intra-individual variability in bile salt concentrations in vivo which may contribute to low and variable oral peptide bioavailability (typically <0.1%). The aim of this study was to investigate the interaction of the model peptide drug, octreotide, with biologically relevant bile salts in solution. In vitro flux studies revealed octreotide to interact with seven different bile salts in solution at monomeric and micellar concentrations. Dihydroxy bile salts had a significantly larger effect on octreotide membrane flux than trihydroxy bile salts.

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Diffusion Order Spectroscopy (DOSY) was used as an NMR-based biophysical screen to establish bile salt micelle size and confirm the association of octreotide with micelles. Chemical shift perturbations of octreotide upon titration of bile salts from below to above the critical micellar concentration established specific loci of intermolecular interactions. High resolution 2D 1H-1H NOESY spectra further dissected these interactions and suggests that the interface between aromatic protons of the peptide and hydrophobic moieties of bile salts drive the association. Ala-mutated octreotide analogs were used to challenge this interface. Overall, these results improve the understanding of peptide drug-bile salt interactions and will aid the development of orally bioavailable peptide drugs.

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13th Great Plains Regional Annual

Symposium on Protein and Biomolecular NMR

Participant’s List and

Contact Information

October 9-10, 2020

University of Kansas

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Last Name First Name Institution Affiliation Email Address Acquaah-Harrison George Milliporesigma ga358404@ohio.edu An Yeongseo Iowa State University yan1@iastate.edu Anderson Austin Oklahoma State University Aaustir@okstate.edu Anklin Clemens Bruker BioSpin clemens.anklin@bruker.com Arens Christina Missouri University of Science and

Technology cmatvc@mst.edu

Atsrim Ernest University of Nebraska-Lincoln eatsrim2@huskers.unl.edu Beck Moriah Wichita State University moriah.beck@wichita.edu Boehmer Steven MilliporeSigma - Isotec steven.boehmer@milliporesigma.com Bryce James Oklahoma State University jr.bryce@okstate.edu Chakravarty Amritangshu The University of Kansas a718c558@ku.edu Cook Gabriel Oklahoma State University gabriel.cook@okstate.edu Cronin Josh MilliporeSigma - Isotec joshua.cronin@milliporesigma.com De Guzman Roberto The University of Kansas rdguzman@ku.edu Dembele Hawa Kansas State University hpdembele@ksu.edu Dening Tahnee The University of Kansas tahneedening@ku.edu Dlamini Nkosinathi University of Nebraska-Lincoln ndlamini2@huskers.unl.edu Douglas Justin The University of Kansas justindouglas@ku.edu Dupureur Cynthia University of Missouri - St. Louis cdup@umsl.edu Eichhorn Catherine University of Nebraska-Lincoln ceichhor@unl.edu Elballa Waleed The University of Kansas welballa@ku.edu Foster Avery University of Missouri alfb28@mail.missouri.edu Gremminger Thomas University of Missouri - Columbia tjgyt7@mail.missouri.edu Guha Biswas Pallavi The University of Kansas pallaviguhabiswas@ku.edu Hageman Michael The University of Kansas mhageman@ku.edu Halimeh Wyel Wichita State University wxhalimeh@shockers.wichita.edu Hall Randal Cambridge Isotope Labs rhall@isotope.com Harris Michael Oklahoma State Universtiy michael.s.harris10@okstate.edu Henen Morkos University of Colorado morkos.henen@cuanschutz.edu Heng Xiao University of Missouri - Columbia hengx@missouri.edu Jafari Negar The University of Kansas Negar.jafari@ku.edu Jarrett Shelby University of Missouri - St. Louis sjarr2@gmail.com Jayasekera Vishakha University of Nebraska-Lincoln udesilvajayasekera2@huskers.unl.edu Ji Juan University of Missouri jij@missouri.edu Johnson Collin Missouri State University collin314@live.missouristate.edu Juncos Juliana Leloir Institute maria.juliana207@gmail.com Khatiwada Balabhadra Iowa State University bal@iastate.edu Klausmeyer Rachel Wichita State University raklausmeyer@shockers.wichita.edu Liu Yijun Iowa State yijunl@iastate.edu Lou Hao The University of Kansas lou0@ku.edu Lowe Jacques Iowa State University jacquesl@iastate.edu

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Last Name First Name Institution Affiliation Email Address Maddali Srikanth Milliporesigma SRI.MADDALI@MILLIPORESIGMA.COM McMullin Cole University of Missouri - Kansas City cpmrhb@umsystem.edu Meeks Kaylen University of Missouri kmcbp@mail.missouri.edu Meints Gary Missouri State University garymeints@missouristate.edu Mercedes Kilsia University of Colorado - Anschutz

Medical School kilsia.mercedes@cuanschutz.edu

Mishra Nitin Kansas State University nitinm@ksu.edu Mohamed Amr University of Nebraska-Lincoln amr.mohamed199815@yahoo.com Nguyen Trang Iowa State University trangnt@iastate.edu Ogunleye Tiwalola Oklahoma State University togunle@okstate.edu Pal Indrani Kansas State University ipal@ksu.edu Pilarski Autumn Missouri State University pilarski4672@live.missouristate.edu Prakash Om Kansas State University omp@ksu.edu Purslow Jeff Iowa State University jpurslow@iastate.edu Reiter Nicholas Marquette University nicholas.reiter@marquette.edu Roche Julien Iowa State University jroche@iastate.edu Schmidt Thomas National Institutes of Health schmidt.nedlitz.ts@gmail.com Shea Madeline University of Iowa madeline-shea@uiowa.edu Singh Aayushi Iowa state University aayushis@iastate.edu Song Zhenwei University of Missouri - Columbia songzh@missouri.edu Spikes Helena University of Missouri - St. Louis hjs7c5@umsystem.edu Staller Seth University of Missouri - Columbia saspn4@umsystem.edu Stieglitz Sarah University of Missouri - Columbia seszkc@umsystem.edu Tabatabai Louisa Iowa State University Lbt@iastate.edu Venditti Vincenzo Iowa State University venditti@iastate.edu Wang George University of Kansas georgewang@ku.edu Weng Iris University of Missouri kwtf8@umsystem.edu Westwood Megan Missouri State University westwood16@live.missouristate.edu Wimberly-Gard Gina CU Anschutz Gina.wimberly-Gard@cuanschutz.edu Wojnarowicz Mark Milliporesigma mark.wojnarowicz@milliporesigma.com Wu Haifan Wichita State University haifan.wu@wichita.edu Yang Kaidi University of Missouri - Columbia kyn69@umsystem.edu Yao Xiaolan University of Missouri - Kansas City yaoxia@umkc.edu Zarena Dr. D. JNTUA CEA zareenajntua@gmail.com Znosko Brent Saint Louis University znoskob@slu.edu

A special Thank You to Nicole Suchy from the Higuchi Biosciences Center for

making GRASP NMR possible

Thank you for attending!