title: Helical alpha-synuclein forms highly conductive ion channels.
abstract: Alpha-synuclein (alphaS) is a cytosolic protein involved in the etiology of Parkinson's disease (PD). Disordered in an aqueous environment, alphaS develops a highly helical conformation when bound to membranes having a negatively charged surface and a large curvature. It exhibits a membrane-permeabilizing activity that has been attributed to oligomeric protofibrillar forms. In this study, monomeric wild-type alphaS and two mutants associated with familial PD, E46K and A53T, formed ion channels with well-defined conductance states in membranes containing 25-50% anionic lipid and 50% phosphatidylethanolamine (PE) in the presence of a trans-negative potential. Another familial mutant, A30P, known to have a lower membrane affinity, did not form ion channels. Ca2+ prevented channel formation when added to membranes before alphaS and decreased channel conductance when added to preformed channels. In contrast to the monomer, membrane permeabilization by oligomeric alphaS was not characterized by formation of discrete channels, a requirement for PE lipid, or a membrane potential. Channel activity, alpha-helical content, thermal stability of membrane-bound alphaS determined by far-UV CD, and lateral mobility of alphaS bound to planar membranes measured by fluorescence correlation spectroscopy were correlated. It was inferred that discrete ion channels with well-defined conductance states were formed in the presence of a membrane potential by one or several molecules of monomeric alphaS in an alpha-helical conformation and that such channels may have a role in the normal function and/or pathophysiology of the protein.
authors.affiliation: Departments of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-2054, USA.
authors.forename: Stanislav D
authors.initials: SD
authors.lastname: Zakharov
authors.forename: John D
authors.initials: JD
authors.lastname: Hulleman
authors.forename: Elena A
authors.initials: EA
authors.lastname: Dutseva
authors.forename: Yuri N
authors.initials: YN
authors.lastname: Antonenko
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Rochet
authors.forename: William A
authors.initials: WA
authors.lastname: Cramer
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D019281:Dimerization; D006801:Humans; D007473:Ion Channels / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D008055:Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D007473:Ion Channels; D008055:Lipids; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D051844:alpha-Synuclein; C017251:1,2-oleoylphosphatidylcholine