Identification of Novel α-Synuclein Isoforms in Human Brain Tissue by using an Online NanoLC-ESI-FTICR-MS Method

• Published in: Neurochemical research Vol. 36; no. 11; pp. 2029 - 2042
• Main Authors: Öhrfelt, Annika, Zetterberg, Henrik, Andersson, Kerstin, Persson, Rita, Secic, Dzemila, Brinkmalm, Gunnar, Wallin, Anders, Mulugeta, Ezra, Francis, Paul T., Vanmechelen, Eugeen, Aarsland, Dag, Ballard, Clive, Blennow, Kaj, Westman-Brinkmalm, Ann
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.11.2011
• Subjects: Aged; Aged, 80 and over; alpha-Synuclein - chemistry; alpha-Synuclein - isolation & purification; Amino Acid Sequence; Biochemistry; Biomedical and Life Sciences; Biomedicine; Blotting, Western; Brain; Brain Chemistry; Brodmann's area; Cell Biology; Cerebrospinal fluid; Cortex (prefrontal); Cortex (temporal); Dementia disorders; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoprecipitation; Internet; Ionization; Lewy bodies; Lewy Body Disease - metabolism; Liquid chromatography; Male; Mass spectrometry; Mass spectroscopy; Medicin och hälsovetenskap; Middle Aged; Movement disorders; Nanotechnology; Neurochemistry; Neurodegenerative diseases; Neurology; Neurosciences; Original Paper; Parkinson Disease - metabolism; Parkinson's disease; Protein Isoforms - chemistry; Protein Isoforms - isolation & purification; Psychiatry; Psykiatri; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Synuclein; Tandem Mass Spectrometry; Western blotting; Immunoprecipitation; Cerebrospinal fluid; Neurodegenerative diseases; Synuclein; Mass spectrometry
• ISSN: 0364-3190; 1573-6903; 1573-6903
• DOI: 10.1007/s11064-011-0527-x

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-syn 1–140 ) and two N-terminally acetylated C-terminally truncated forms of α-syn (Ac-α-syn 1–139 and Ac-α-syn 1–103 ) 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).