Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells
description
Transcript of Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells
![Page 1: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/1.jpg)
Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells
Chuan HeUniversity of Chicago
Department of Chemistry
Mark P. JensenArgonne National LabChemical Sciences and Engineering Division
Sixth Argonne – UChicago – Fermilab Collaboration Meeting, October 12, 2009
![Page 2: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/2.jpg)
2
The Nuclear Conundrum
Nuclear Energy is a carbon-free source for 20% of U.S. & world electricity production.
Why not 80%?
– Socio-economic reasons• Cost and economic risk• Proliferation of nuclear technology• Public worries about safety
– Technical reason• How do we handle the waste?
![Page 3: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/3.jpg)
3
Actinides? Really?
Recognizing metal ions with high selectivity and sensitivity is essential for life.Nature has evolved various metalloproteins to exploit and control metals.
![Page 4: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/4.jpg)
4
Objectives
Discover how cells handle toxic human-made metals, the transuranium actinides
1. Visualize actinide trafficking in cells with X-ray microbeams
2. Identify individual proteins that bind actinides in mammalian cells
3. Biochemically and structurally characterize actinide binding by various proteins
Create new ways to efficiently, economically, and safelyhandle actinides when they are no longer useful
![Page 5: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/5.jpg)
5
Where do Actinides Accumulate in Cells?
![Page 6: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/6.jpg)
6
First Structures of Pu-Containing Proteins
Small Angle X-Ray Scattering of Transferrins
One form of transferrin containingboth plutonium and iron has thesame shape as the native protein,which contains only iron.
![Page 7: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/7.jpg)
7
Blocking Pu Uptake by Cells Appears Possible
No Plutonium
“Bad” PlutoniumPuCFeNTf
“Bad” Plutoniumplus 50 uMchloroquine
Images of rat adrenal gland cellsAxes - coordinates in mm
![Page 8: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/8.jpg)
8
Could metal-binding proteins be redesigned to bind actinides?
NikR is a transcriptional repressor for expression of the nikABCDE operon in the presence of excessive concentrations of intracellular Ni+2
Ref: C. L. Drennan et al, Nat. Struc. Biol., 2003, 10, 794-799C. L. Drennan et al, PNAS, 2006, 103, 13676-13681
NikR
![Page 9: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/9.jpg)
9
MQRVTITLDD DLLETLDSLS QRRGYNNRSE AIRDILRSAL AQEATQQHGT QGFAVLSYVY EHEKRDLASR IVSTQHHHHD LSVATLHVHI NHDDCLEIAV LKGDMGDVQH FADDVIAQRG VRHGHLQCLP KED
Mutations:V72 → S V72SC95→ D, H, S V72S C95D H76→ D,E
V72S C95S H76→ D,E
NiN
S
N
NN
NHN
H
H
O NH
V72
3.7 A
4.4 AH76'
C95
H89
H87
UN
S
N
NN
NHN
H
H
O
O NH
V72S
H76'
C95
H89
H87
H
O
O
V72SU
N
NN
N
H
H
O
O NH
V72S
C95D
H89
H87
H
O
O
OO
OO
H76'D
H76D C95D
Design of an Actinyl Binding Site
![Page 10: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/10.jpg)
10
Design of an Actinyl Binding Site
The Kd of uranyl binding to NikR’was determined to be 53 nM
![Page 11: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/11.jpg)
11
Uranyl Binding and Function
![Page 12: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/12.jpg)
12
S. V. Wegner, H. Boyaci, H. Chen, M. P. Jensen, C. He, Angew. Chem. Int. Ed. (2009), 48, 2339.
Joint Publication
![Page 13: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/13.jpg)
13
Engineer bacteria to uptake, transport and store actinides
![Page 14: Elucidation and Control of Actinide Trafficking Pathways in Mammalian Cells](https://reader035.fdocuments.us/reader035/viewer/2022062322/56814748550346895db48654/html5/thumbnails/14.jpg)
14
Design Peptides that are Selective for Specific Actinides
LBT: DTNNDGWYEGDELLA1 53 9 12