title: Deamidation of alpha-synuclein.
abstract: The rates of deamidation of alpha-synuclein and single Asn residues in 13 Asn-sequence mutants have been measured for 5 x 10(-5)M protein in both the absence and presence of 10(-2)M sodium dodecyl sulfate (SDS). In the course of these experiments, 370 quantitative protein deamidation measurements were performed and 37 deamidation rates were determined by ion cyclotron resonance Fourier transform mass spectrometry, using an improved whole protein isotopic envelope method and a mass defect method with both enzymatic and collision-induced fragmentation. The measured deamidation index of alpha-synuclein was found to be 0.23 for an overall deamidation half-time of 23 days, without or with SDS micelles, owing primarily to the deamidation of Asn(103) and Asn(122). Deamidation rates of 15 Asn residues in the wild-type and mutant proteins were found to be primary sequence controlled without SDS. However, the presence of SDS micelles slowed the deamidation rates of nine N-terminal region Asn residues, caused by the known three-dimensional structures induced through protein binding to SDS micelles.
authors.affiliation: Oregon Institute of Science and Medicine, Cave Junction, Oregon 97523, USA. noah@oism.org
authors.forename: Noah E
authors.initials: NE
authors.lastname: Robinson
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Robinson
authors.forename: Stephanie E S
authors.initials: SE
authors.lastname: Schulze
authors.forename: Bert T
authors.initials: BT
authors.lastname: Lai
authors.forename: Harry B
authors.initials: HB
authors.lastname: Gray
mesh_terms: D000577:Amides / Q000737:chemistry*; D000595:Amino Acid Sequence; D001216:Asparagine / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics
chemical_list: D000577:Amides; D011994:Recombinant Proteins; D051844:alpha-Synuclein; D012967:Sodium Dodecyl Sulfate; D001216:Asparagine