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A protease for 'middle-down' proteomics

Orbitrap_SciLib
Reputable Mentor II
Reputable Mentor II
Wu C, Tran JC, Zamdborg L, Durbin KR, Li M, Ahlf DR, Early BP, Thomas PM, Sweedler JV, Kelleher NL.
Nat Methods. 2012 Jun 17;9(8):822-4.
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.

http://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.2074.html
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
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