title: Identification of novel α-synuclein isoforms in human brain tissue by using an online nanoLC-ESI-FTICR-MS method.
abstract: Parkinson's disease (PD) and Dementia with Lewy bodies (DLB) are neurodegenerative diseases that are characterized by intra-neuronal inclusions of Lewy bodies in distinct brain regions. These inclusions consist mainly of aggregated α-synuclein (α-syn) protein. The present study used immunoprecipitation combined with nanoflow liquid chromatography (LC) coupled to high resolution electrospray ionization Fourier transform ion cyclotron resonance tandem mass spectrometry (ESI-FTICR-MS/MS) to determine known and novel isoforms of α-syn in brain tissue homogenates. N-terminally acetylated full-length α-syn (Ac-α-syn₁₋₁₄₀) and two N-terminally acetylated C-terminally truncated forms of α-syn (Ac-α-syn₁₋₁₃₉ and Ac-α-syn₁₋₁₀₃) were found. The different forms of α-syn were further studied by Western blotting in brain tissue homogenates from the temporal cortex Brodmann area 36 (BA36) and the dorsolateral prefrontal cortex BA9 derived from controls, patients with DLB and PD with dementia (PDD). Quantification of α-syn in each brain tissue fraction was performed using a novel enzyme-linked immunosorbent assay (ELISA).
authors.affiliation: Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, S-43180 Mölndal, Sweden. annika.ohrfelt@neuro.gu.se
authors.forename: Annika
authors.initials: A
authors.lastname: Ohrfelt
authors.forename: Henrik
authors.initials: H
authors.lastname: Zetterberg
authors.forename: Kerstin
authors.initials: K
authors.lastname: Andersson
authors.forename: Rita
authors.initials: R
authors.lastname: Persson
authors.forename: Dzemila
authors.initials: D
authors.lastname: Secic
authors.forename: Gunnar
authors.initials: G
authors.lastname: Brinkmalm
authors.forename: Anders
authors.initials: A
authors.lastname: Wallin
authors.forename: Ezra
authors.initials: E
authors.lastname: Mulugeta
authors.forename: Paul T
authors.initials: PT
authors.lastname: Francis
authors.forename: Eugeen
authors.initials: E
authors.lastname: Vanmechelen
authors.forename: Dag
authors.initials: D
authors.lastname: Aarsland
authors.forename: Clive
authors.initials: C
authors.lastname: Ballard
authors.forename: Kaj
authors.initials: K
authors.lastname: Blennow
authors.forename: Ann
authors.initials: A
authors.lastname: Westman-Brinkmalm
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D015153:Blotting, Western; D001923:Brain Chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020961:Lewy Body Disease / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D036103:Nanotechnology; D010300:Parkinson Disease / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D051844:alpha-Synuclein / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D020033:Protein Isoforms; D051844:alpha-Synuclein