Lipid Dynamics and Protein–Lipid Interactions in 2D Crystals Formed with the β-Barrel Integral Membrane Protein VDAC1
Author(s)Ong, Ta-Chung; Clark, Lindsay; Teijido, Oscar; van der Wel, Patrick C. A.; Garces, Robert; Wagner, Gerhard; Rostovtseva, Tatiana K.; Eddy, Matthew Thomas; Griffin, Robert Guy; ... Show more Show less
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We employ a combination of [superscript 13]C/[superscript 15]N magic angle spinning (MAS) NMR and [superscript 2]H NMR to study the structural and functional consequences of different membrane environments on VDAC1 and, conversely, the effect of VDAC1 on the structure of the lipid bilayer. MAS spectra reveal a well-structured VDAC1 in 2D crystals of dimyristoylphosphatidylcholine (DMPC) and diphytanoylphosphatidylcholine (DPhPC), and their temperature dependence suggests that the VDAC structure does not change conformation above and below the lipid phase transition temperature. The same data show that the N-terminus remains structured at both low and high temperatures. Importantly, functional studies based on electrophysiological measurements on these same samples show fully functional channels, even without the presence of Triton X-100 that has been found necessary for in vitro-refolded channels. [superscript 2]H solid-state NMR and differential scanning calorimetry were used to investigate the dynamics and phase behavior of the lipids within the VDAC1 2D crystals. [superscript 2]H NMR spectra indicate that the presence of protein in DMPC results in a broad lipid phase transition that is shifted from 19 to ~27 °C and show the existence of different lipid populations, consistent with the presence of both annular and bulk lipids in the functionally and structurally homogeneous samples.
DepartmentMassachusetts Institute of Technology. Department of Chemistry; Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
Journal of the American Chemical Society
American Chemical Society (ACS)
Eddy, Matthew T., Ta-Chung Ong, Lindsay Clark, Oscar Teijido, Patrick C. A. van der Wel, Robert Garces, Gerhard Wagner, Tatiana K. Rostovtseva, and Robert G. Griffin. “Lipid Dynamics and Protein–Lipid Interactions in 2D Crystals Formed with the β-Barrel Integral Membrane Protein VDAC1.” Journal of the American Chemical Society 134, no. 14 (April 11, 2012): 6375-6387.
Author's final manuscript