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

doi:10.1038/nmeth.2074

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 journal › Article

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 language	English (US)
Pages (from-to)	822-824
Number of pages	3
Journal	Nature Methods
Volume	9
Issue number	8
DOIs	https://doi.org/10.1038/nmeth.2074
State	Published - Aug 1 2012

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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Wu, C., Tran, J. C., Zamdborg, L., Durbin, K. R., Li, M., Ahlf, D. R., Early, B. P., Thomas, P. M., Sweedler, J. V., & Kelleher, N. L. (2012). A protease for 'middle-down' proteomics. Nature Methods, 9(8), 822-824. https://doi.org/10.1038/nmeth.2074

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AU - Ahlf, Dorothy R.

AU - Early, Bryan P.

AU - Thomas, Paul M.

AU - Sweedler, Jonathan V.

AU - Kelleher, Neil L.

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