Robust analysis of the yeast proteome under 50 kDa by molecular-mass-based fractionation and top-down mass spectrometry

John F. Kellie, Adam D. Catherman, Kenneth R. Durbin, John C. Tran, Jeremiah D. Tipton, Jeremy L. Norris, Charles E. Witkowski, Paul Martin Thomas, Neil L Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

As the process of top-down mass spectrometry continues to mature, we benchmark the next installment of an improving methodology that incorporates a tube-gel electrophoresis (TGE) device to separate intact proteins by molecular mass. Top-down proteomics is accomplished in a robust fashion to yield the identification of hundreds of unique proteins, many of which correspond to multiple protein forms. The TGE platform separates 0-50 kDa proteins extracted from the yeast proteome into 12 fractions prior to automated nanocapillary LC-MS/MS in technical triplicate. The process may be completed in less than 72 h. From this study, 530 unique proteins and 1103 distinct protein species were identified and characterized, thus representing the highest coverage to date of the Saccharomyces cerevisiae proteome using top-down proteomics. The work signifies a significant step in the maturation of proteomics based on direct measurement and fragmentation of intact proteins.

Original languageEnglish (US)
Pages (from-to)209-215
Number of pages7
JournalAnalytical Chemistry
Volume84
Issue number1
DOIs
StatePublished - Jan 3 2012

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint Dive into the research topics of 'Robust analysis of the yeast proteome under 50 kDa by molecular-mass-based fractionation and top-down mass spectrometry'. Together they form a unique fingerprint.

  • Cite this