A protease for 'middle-down' proteomics

Cong Wu, John C. Tran, Leonid Zamdborg, Kenneth R. Durbin, Mingxi Li, Dorothy R. Ahlf, Bryan P. Early, Paul M. Thomas, Jonathan V. Sweedler, Neil L. Kelleher

Research output: Contribution to journalArticlepeer-review

Abstract

We developed a method for restricted enzymatic proteolysis using the outer membrane protease T (OmpT) to produce large peptides (>6.3 kDa on average) for mass spectrometry-based proteomics. Using this approach to analyze prefractionated high-mass HeLa proteins, we identified 3,697 unique peptides from 1,038 proteins. We demonstrated the ability of large OmpT peptides to differentiate closely related protein isoforms and to enable the detection of many post-translational modifications.

Original languageEnglish (US)
Pages (from-to)822-824
Number of pages3
JournalNature Methods
Volume9
Issue number8
DOIs
StatePublished - Aug 2012

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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