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  Structure and gating behavior of the human integral membrane protein VDAC1 in a lipid bilayer

Najbauer, E. E., Tekwani Movellan, K., Giller, K., Benz, R., Becker, S., Griesinger, C., et al. (2022). Structure and gating behavior of the human integral membrane protein VDAC1 in a lipid bilayer. Journal of the American Chemical Society, 144(7), 2953-2967. doi:10.1021/jacs.1c09848.

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 Creators:
Najbauer, E. E.1, Author           
Tekwani Movellan, K.1, Author           
Giller, K.2, Author           
Benz, R., Author
Becker, S.2, Author           
Griesinger, C.2, Author                 
Andreas, L. B.1, Author           
Affiliations:
1Research Group of Solid State NMR Spectroscopy-2, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350125              
2Department of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350124              

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Free keywords: solid-state nmr; dependent anion channel; permeability transition pore; beta-barrel; backbone assignment; correlation spectroscopy; ;selective channel; binding-site; resolution; reconstitution
 Abstract: The voltage-dependent anion channel (VDAC), the most abundant protein in the outer mitochondrial membrane, is responsible for the transport of all ions and metabolites into and out of mitochondria. Larger than any of the beta-barrel structures determined to date by magic-angle spinning (MAS) NMR, but smaller than the size limit of cryo-electron microscopy (cryo-EM), VDAC1's 31 kDa size has long been a bottleneck in determining its structure in a near-native lipid bilayer environment. Using a single two-dimensional (2D) crystalline sample of human VDAC1 in lipids, we applied proton-detected fast magic-angle spinning NMR spectroscopy to determine the arrangement of beta strands. Combining these data with long-range restraints from a spin-labeled sample, chemical shift-based secondary structure prediction, and previous MAS NMR and atomic force microscopy (AFM) data, we determined the channel's structure at a 2.2 angstrom root-mean-square deviation (RMSD). The structure, a 19-stranded beta-barrel, with an N-terminal alpha-helix in the pore is in agreement with previous data in detergent, which was questioned due to the potential for the detergent to perturb the protein's functional structure. Using a quintuple mutant implementing the channel's closed state, we found that dynamics are a key element in the protein's gating behavior, as channel closure leads to the destabilization of not only the C-terminal barrel residues but also the alpha 2 helix. We showed that cholesterol, previously shown to reduce the frequency of channel closure, stabilizes the barrel relative to the N-terminal helix. Furthermore, we observed channel closure through steric blockage by a drug shown to selectively bind to the channel, the Bcl2-antisense oligonucleotide G3139.

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Language(s): eng - English
 Dates: 2022-02-142022-02-23
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1021/jacs.1c09848
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Project name : SFB 803
Grant ID : -
Funding program : (project A04)
Funding organization : DFG
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Grant ID : AN1316/1-1
Funding program : Emmy Noether Program
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Funding organization : Max Planck Society

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Title: Journal of the American Chemical Society
Source Genre: Journal
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Pages: - Volume / Issue: 144 (7) Sequence Number: - Start / End Page: 2953 - 2967 Identifier: ISSN: 0002-7863
ISSN: 1520-5126