title: Quantitative measurement of intact alpha-synuclein proteoforms from post-mortem control and Parkinson's disease brain tissue by intact protein mass spectrometry.
abstract: A robust top down proteomics method is presented for profiling alpha-synuclein species from autopsied human frontal cortex brain tissue from Parkinson's cases and controls. The method was used to test the hypothesis that pathology associated brain tissue will have a different profile of post-translationally modified alpha-synuclein than the control samples. Validation of the sample processing steps, mass spectrometry based measurements, and data processing steps were performed. The intact protein quantitation method features extraction and integration of m/z data from each charge state of a detected alpha-synuclein species and fitting of the data to a simple linear model which accounts for concentration and charge state variability. The quantitation method was validated with serial dilutions of intact protein standards. Using the method on the human brain samples, several previously unreported modifications in alpha-synuclein were identified. Low levels of phosphorylated alpha synuclein were detected in brain tissue fractions enriched for Lewy body pathology and were marginally significant between PD cases and controls (p = 0.03).
authors: GSK, 709 Swedeland Road, King of Prussia, PA.
authors: John F
authors: JF
authors: Kellie
authors: Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN.
authors: Richard E
authors: RE
authors: Higgs
authors: Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN.
authors: John W
authors: JW
authors: Ryder
authors: Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN.
authors: Anthony
authors: A
authors: Major
authors: Civin Laboratory for Neuropathology, Banner Sun Health Research Institute, Sun City, AZ.
authors: Thomas G
authors: TG
authors: Beach
authors: Department of Neurology, Mayo Clinic Arizona, Scottsdale, AZ.
authors: Charles H
authors: CH
authors: Adler
authors: TransThera Consulting Co., Indianapolis, IN.
authors: Kalpana
authors: K
authors: Merchant
authors: Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN.
authors: Michael D
authors: MD
authors: Knierman
mesh_terms: D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002102:Cadaver; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D003627:Data Interpretation, Statistical; D005260:Female; D006801:Humans; D010300:Parkinson Disease / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D051844:alpha-Synuclein / Q000032:analysis* / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D051844:alpha-Synuclein