title: Role of individual methionines in the fibrillation of methionine-oxidized alpha-synuclein.
abstract: The aggregation of normally soluble alpha-synuclein in the dopaminergic neurons of the substantia nigra is a crucial step in the pathogenesis of Parkinson's disease. Oxidative stress is believed to be a contributing factor in this disorder. We have previously established that oxidation of all four methionine residues in alpha-synuclein (to the sulfoxide, MetO) inhibits fibrillation of this protein in vitro and that the MetO protein also inhibits fibrillation of unmodified alpha-synuclein. Here we show that the degree of inhibition of fibrillation by MetO alpha-synuclein is proportional to the number of oxidized methionines. This was accomplished be selectively converting Met residues into Leu, prior to Met oxidation. The results showed that with one oxidized Met the kinetics of fibrillation were comparable to those for the control (nonoxidized), and with increasing numbers of methionine sulfoxides the kinetics of fibrillation became progressively slower. Electron microscope images showed that the fibril morphology was similar for all species examined, although fewer fibrils were observed with the oxidized forms. The presence of zinc was shown to overcome the Met oxidation-induced inhibition. Interestingly, substitution of Met by Leu led to increased propensity for aggregation (soluble oligomers) but slower formation of fibrils.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95064, USA.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Hokenson
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
authors.forename: John
authors.initials: J
authors.lastname: Goers
authors.forename: Ghiam
authors.initials: G
authors.lastname: Yamin
authors.forename: Larissa A
authors.initials: LA
authors.lastname: Munishkina
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Fink
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D002942:Circular Dichroism; D005454:Fluorescence Polarization; D006801:Humans; D007930:Leucine / Q000235:genetics; D008027:Light; D008715:Methionine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D017550:Spectroscopy, Fourier Transform Infrared; D051843:Synucleins; D015032:Zinc / Q000737:chemistry; D051844:alpha-Synuclein
chemical_list: D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; D051843:Synucleins; D051844:alpha-Synuclein; D008715:Methionine; D007930:Leucine; D015032:Zinc; C013111:methionine sulfoxide