title: Structural and functional characterization of the integral membrane protein VDAC-1 in lipid bilayer nanodiscs.
abstract: Biophysical studies of membrane proteins are often impeded by the requirement for a membrane mimicking environment. Detergent micelles are the most common choice, but the denaturing properties make them unsatisfactory for studies of many membrane proteins and their interactions. In the present work, we explore phospholipid bilayer nanodiscs as membrane mimics and employ electron microscopy and solution NMR spectroscopy to characterize the structure and function of the human voltage dependent anion channel (VDAC-1) as an example of a polytopic integral membrane protein. Electron microscopy reveals the formation of VDAC-1 multimers, an observation that is consistent with results obtained in native mitochondrial outer membranes. High-resolution NMR spectroscopy demonstrates a well folded VDAC-1 protein and native NADH binding functionality. The observed chemical shift changes upon addition of the native ligand NADH to nanodisc-embedded VDAC-1 resemble those of micelle-embedded VDAC-1, indicating a similar structure and function in the two membrane-mimicking environments. Overall, the ability to study integral membrane proteins at atomic resolution with solution NMR in phospholipid bilayers, rather than in detergent micelles, offers exciting novel possibilities to approach the biophysical properties of membrane proteins under nondenaturing conditions, which makes this technology particular suitable for protein-protein interactions and other functional studies.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Raschle
authors.forename: Sebastian
authors.initials: S
authors.lastname: Hiller
authors.forename: Tsyr-Yan
authors.initials: TY
authors.lastname: Yu
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Rice
authors.forename: Thomas
authors.initials: T
authors.lastname: Walz
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wagner
mesh_terms: D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D011487:Protein Conformation; D013499:Surface Properties; D050995:Voltage-Dependent Anion Channel 1 / Q000737:chemistry*
chemical_list: D008051:Lipid Bilayers; C494762:VDAC1 protein, human; D050995:Voltage-Dependent Anion Channel 1; D004134:Dimyristoylphosphatidylcholine