A protease for 'middledown' proteomics

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...

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Bibliographic Details
Published inNature methods Vol. 9; no. 8; p. 822
Main Authors Wu, Cong, Tran, John C, Zamdborg, Leonid, Durbin, Kenneth R, Li, Mingxi, Ahlf, Dorothy R, Early, Bryan P, Thomas, Paul M, Sweedler, Jonathan V, Kelleher, Neil L
Format Journal Article
LanguageEnglish
Published Nature Publishing Group 01.08.2012
Subjects
Mass spectrometry
Physiological aspects
Proteases
Proteolysis
United States
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Summary: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.
ISSN:1548-7091
1548-7105
DOI:10.1038/NMETH.2074