Liquid Chromatography-Tandem Mass Spectrometry to Define Sortase Cleavage Products.
Sortase enzymes have specific endopeptidase activity, cleaving within a defined pentapeptide sequence at the C-terminal end of their protein substrates. Here, we describe how monitoring sortase cleavage activity can be achieved using peptide substrates. Peptide cleavage can be readily analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS), which allows for the precise definition of cleavage sites. This technique could be used to analyze the peptidase activity of any enzyme, and identify sites of cleavage within any peptide.
Department of Biology, McMaster University, Hamilton, ON, Canada.|Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
Andrew
A
Duong
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.|Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada.
Kalinka
K
Koteva
Department of Biology, McMaster University, Hamilton, ON, Canada.|Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
Danielle L
DL
Sexton
Department of Biology, McMaster University, Hamilton, ON, Canada. melliot@mcmaster.ca.|Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Life Science Building, RM 329, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada. melliot@mcmaster.ca.
Marie A
MA
Elliot
D002853:Chromatography, Liquid / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D059748:Proteolysis; D053719:Tandem Mass Spectrometry / Q000379:methods*
D010455:Peptides; D003546:Cysteine Endopeptidases
Liquid chromatography; Mass spectrometry; Peptidase; Peptide cleavage; Sortase