title: Phosphorylation of Ser-129 is the dominant pathological modification of alpha-synuclein in familial and sporadic Lewy body disease.
abstract: A comprehensive, unbiased inventory of synuclein forms present in Lewy bodies from patients with dementia with Lewy bodies was carried out using two-dimensional immunoblot analysis, novel sandwich enzyme-linked immunosorbent assays with modification-specific synuclein antibodies, and mass spectroscopy. The predominant modification of alpha-synuclein in Lewy bodies is a single phosphorylation at Ser-129. In addition, there is a set of characteristic modifications that are present to a lesser extent, including ubiquitination at Lys residues 12, 21, and 23 and specific truncations at Asp-115, Asp-119, Asn-122, Tyr-133, and Asp-135. No other modifications are detectable by tandem mass spectrometry mapping, except for a ubiquitous N-terminal acetylation. Small amounts of Ser-129 phosphorylated and Asp-119-truncated alpha-synuclein are present in the soluble fraction of both normal and disease brains, suggesting that these Lewy body-associated forms are produced during normal metabolism of alpha-synuclein. In contrast, ubiquitination is only detected in Lewy bodies and is primarily present on phosphorylated synuclein; it therefore likely occurs after phosphorylated synuclein has deposited into Lewy bodies. This invariant pattern of specific phosphorylation, truncation, and ubiquitination is also present in the detergent-insoluble fraction of brain from patients with familial Parkinson's disease (synuclein A53T mutation) as well as multiple system atrophy, suggesting a common pathogenic pathway for both genetic and sporadic Lewy body diseases. These observations are most consistent with a model in which preferential accumulation of normally produced Ser-129 phosphorylated alpha-synuclein is the key event responsible for the formation of Lewy bodies in various Lewy body diseases.
authors.affiliation: Elan Pharmaceuticals, South San Francisco, California 94080, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Anderson
authors.forename: Donald E
authors.initials: DE
authors.lastname: Walker
authors.forename: Jason M
authors.initials: JM
authors.lastname: Goldstein
authors.forename: Rian
authors.initials: R
authors.lastname: de Laat
authors.forename: Kelly
authors.initials: K
authors.lastname: Banducci
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Caccavello
authors.forename: Robin
authors.initials: R
authors.lastname: Barbour
authors.forename: Jiping
authors.initials: J
authors.lastname: Huang
authors.forename: Kristin
authors.initials: K
authors.lastname: Kling
authors.forename: Michael
authors.initials: M
authors.lastname: Lee
authors.forename: Linnea
authors.initials: L
authors.lastname: Diep
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Keim
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Shen
authors.forename: Tim
authors.initials: T
authors.lastname: Chataway
authors.forename: Michael G
authors.initials: MG
authors.lastname: Schlossmacher
authors.forename: Peter
authors.initials: P
authors.lastname: Seubert
authors.forename: Dale
authors.initials: D
authors.lastname: Schenk
authors.forename: Sukanto
authors.initials: S
authors.lastname: Sinha
authors.forename: Wei Ping
authors.initials: WP
authors.lastname: Gai
authors.forename: Tamie J
authors.initials: TJ
authors.lastname: Chilcote
mesh_terms: D000595:Amino Acid Sequence; D005260:Female; D006801:Humans; D020961:Lewy Body Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D008875:Middle Aged; D008969:Molecular Sequence Data; D010766:Phosphorylation; D012694:Serine / Q000235:genetics / Q000378:metabolism*; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein; D012694:Serine