A protease for 'middle-down' 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 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.
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Cong
C
Wu
John C
JC
Tran
Leonid
L
Zamdborg
Kenneth R
KR
Durbin
Mingxi
M
Li
Dorothy R
DR
Ahlf
Bryan P
BP
Early
Paul M
PM
Thomas
Jonathan V
JV
Sweedler
Neil L
NL
Kelleher
D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism*
D010455:Peptides; D020033:Protein Isoforms; D012697:Serine Endopeptidases; C057840:omptin outer membrane protease