Quantitative measurement of intact alpha-synuclein proteoforms from post-mortem control and parkinson's disease brain tissue by intact protein mass spectrometry

John F. Kellie, Richard E. Higgs, John W. Ryder, Anthony Major, Thomas G. Beach, Charles Howard Adler, Kalpana Merchant, Michael D. Knierman

Research output: Contribution to journalArticle

47 Citations (Scopus)

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).

Original languageEnglish (US)
Article number5797
JournalScientific Reports
Volume4
DOIs
StatePublished - Jul 23 2014

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alpha-Synuclein
Parkinson Disease
Mass Spectrometry
Brain
Proteins
Pathology
Lewy Bodies
Frontal Lobe
Proteomics
Linear Models

ASJC Scopus subject areas

  • General

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Quantitative measurement of intact alpha-synuclein proteoforms from post-mortem control and parkinson's disease brain tissue by intact protein mass spectrometry. / Kellie, John F.; Higgs, Richard E.; Ryder, John W.; Major, Anthony; Beach, Thomas G.; Adler, Charles Howard; Merchant, Kalpana; Knierman, Michael D.

In: Scientific Reports, Vol. 4, 5797, 23.07.2014.

Research output: Contribution to journalArticle

Kellie, John F. ; Higgs, Richard E. ; Ryder, John W. ; Major, Anthony ; Beach, Thomas G. ; Adler, Charles Howard ; Merchant, Kalpana ; Knierman, Michael D. / Quantitative measurement of intact alpha-synuclein proteoforms from post-mortem control and parkinson's disease brain tissue by intact protein mass spectrometry. In: Scientific Reports. 2014 ; Vol. 4.
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