title: SNAP-25 contains non-acylated thiol pairs that can form intrachain disulfide bonds: possible sites for redox modulation of neurotransmission.
abstract: Intrachain disulfide bond formation among the cysteine thiols of SNAP-25, a component of the SNARE protein complex required for neurotransmitter release, has been hypothesized to link oxidative stress and inhibition of synaptic transmission. However, neither the availability in vivo of SNAP-25 thiols, which are known targets of S-palmitoylation, nor the tendency of these thiols to form intrachain disulfide bonds is known. We have examined, in rat brain extracts, both the availability of closely spaced, or vicinal, thiol pairs in SNAP-25 and the propensity of these dithiols toward disulfide bond formation using a method improved by us recently that exploits the high chemoselectivity of phenylarsine oxide (PAO) for vicinal thiols. The results show for the first time that a substantial fraction of soluble and, to a lesser extent, particulate SNAP-25 contain non-acylated PAO-binding thiol pairs and that these thiols in soluble SNAP-25 in particular have a high propensity toward disulfide bond formation. Indeed, disulfide bonds were detected in a small fraction of soluble SNAP-25 even under conditions designed to prevent or greatly limit protein thiol oxidation during experimental procedures. These results provide direct experimental support for the availability, in a subpopulation of SNAP-25, of vicinal thiols that may confer on one or more isoforms of this family of proteins a sensitivity to oxidative stress.
authors.affiliation: Departments of Chemistry and Program in Biochemistry, Cell, and Molecular Biology, University of Scranton, Scranton, PA 18510, USA. foleyt2@scranton.edu
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Foley
authors.forename: Abbe R
authors.initials: AR
authors.lastname: Clark
authors.forename: Edward S
authors.initials: ES
authors.lastname: Stredny
authors.forename: Bradley M
authors.initials: BM
authors.lastname: Wierbowski
mesh_terms: D000215:Acylation; D000818:Animals; D001152:Arsenicals / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D012995:Solubility; D013347:Subcellular Fractions / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D009435:Synaptic Transmission / Q000502:physiology*; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism*
chemical_list: D001152:Arsenicals; D004220:Disulfides; C494210:Snap25 protein, rat; D013438:Sulfhydryl Compounds; D050825:Synaptosomal-Associated Protein 25; C029341:oxophenylarsine