ExteNDing Proteome Coverage with Legumain as a Highly Specific Digestion Protease.

ABSTRACT: Bottom-up mass spectrometry-based proteomics utilizes proteolytic enzymes with well characterized specificities to generate peptides amenable for identification by high-throughput tandem mass spectrometry. Trypsin, which cuts specifically after the basic residues lysine and arginine, is the predominant enzyme used for proteome digestion, although proteases with alternative specificities are required to detect sequences that are not accessible after tryptic digest. Here, we show that the human cysteine protease legumain exhibits a strict substrate specificity for cleavage after asparagine and aspartic acid residues during in-solution digestions of proteomes extracted from Escherichia coli, mouse embryonic fibroblast cell cultures, and Arabidopsis thaliana leaves. Generating pe

SUBMITTER: Soh WT 

PROVIDER: S-EPMC7075662 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature