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 › peer-review

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

Access to Document

10.1038/nmeth.2074

Fingerprint Dive into the research topics of 'A protease for 'middle-down' proteomics'. Together they form a unique fingerprint.

Cite this

@article{783080e3d5e54923a5984d8b0a1c08db,

title = "A protease for 'middle-down' proteomics",

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.",

author = "Cong Wu and Tran, {John C.} and Leonid Zamdborg and Durbin, {Kenneth R.} and Mingxi Li and Ahlf, {Dorothy R.} and Early, {Bryan P.} and Thomas, {Paul M.} and Sweedler, {Jonathan V.} and Kelleher, {Neil L.}",

year = "2012",

month = aug,

day = "1",

doi = "10.1038/nmeth.2074",

language = "English (US)",

volume = "9",

pages = "822--824",

journal = "Molecular Pharmaceutics",

issn = "1543-8384",

publisher = "American Chemical Society",

number = "8",

}

TY - JOUR

T1 - A protease for 'middle-down' proteomics

AU - Wu, Cong

AU - Tran, John C.

AU - Zamdborg, Leonid

AU - Durbin, Kenneth R.

AU - Li, Mingxi

AU - Ahlf, Dorothy R.

AU - Early, Bryan P.

AU - Thomas, Paul M.

AU - Sweedler, Jonathan V.

AU - Kelleher, Neil L.

PY - 2012/8/1

Y1 - 2012/8/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84864455743&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864455743&partnerID=8YFLogxK

U2 - 10.1038/nmeth.2074

DO - 10.1038/nmeth.2074

M3 - Article

C2 - 22706673

AN - SCOPUS:84864455743

VL - 9

SP - 822

EP - 824

JO - Molecular Pharmaceutics

JF - Molecular Pharmaceutics

SN - 1543-8384

IS - 8

ER -