A Protease for 'middle-down' Proteomics

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2012 Jun 19

PMID 22706673

Citations 42

Authors

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

Affiliations

Soon will be listed here.

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.

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