title: Solution NMR spectroscopic characterization of human VDAC-2 in detergent micelles and lipid bilayer nanodiscs.
abstract: Three isoforms of the human voltage-dependent anion channel (VDAC), located in the outer mitochondrial membrane, are crucial regulators of mitochondrial function. Numerous studies have been carried out to elucidate biochemical properties, as well as the three-dimensional structure of VDAC-1. However, functional and structural studies of VDAC-2 and VDAC-3 at atomic resolution are still scarce. VDAC-2 is highly similar to VDAC-1 in amino acid sequence, but has substantially different biochemical functions and expression profiles. Here, we report the reconstitution of functional VDAC-2 in lauryldimethylamine-oxide (LDAO) detergent micelles and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayer nanodiscs. We find that VDAC-2 is properly folded in both membrane-mimicking systems and that structural and functional characterization by solution NMR spectroscopy is feasible. This article is part of a Special Issue entitled: VDAC structure, function, and regulation of mitochondrial metabolism.
authors: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors: Tsyr-Yan
authors: TY
authors: Yu
authors: Thomas
authors: T
authors: Raschle
authors: Sebastian
authors: S
authors: Hiller
authors: Gerhard
authors: G
authors: Wagner
mesh_terms: D000595:Amino Acid Sequence; D003902:Detergents / Q000737:chemistry*; D004123:Dimethylamines / Q000737:chemistry; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008823:Micelles; D009243:NAD / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry*; D017510:Protein Folding; D055550:Protein Stability; D012996:Solutions; D013696:Temperature; D050995:Voltage-Dependent Anion Channel 1; D050996:Voltage-Dependent Anion Channel 2 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D003902:Detergents; D004123:Dimethylamines; D008051:Lipid Bilayers; D008823:Micelles; D012996:Solutions; C494762:VDAC1 protein, human; C494765:VDAC2 protein, human; D050996:Voltage-Dependent Anion Channel 2; D009243:NAD; C014518:dodecyldimethylamine oxide; D050995:Voltage-Dependent Anion Channel 1; D004134:Dimyristoylphosphatidylcholine