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- Title
A protease for 'middle-down' proteomics.
- 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
- 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.
- Subjects
PROTEOMICS; PROTEOLYTIC enzymes; PEPTIDES; MASS spectrometry; HELA cells; PROTEINS; POST-translational modification
- Publication
Nature Methods, 2012, Vol 9, Issue 8, p822
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth.2074