Protease Substrate Profiling by N-Terminal COFRADIC.
Detection of (neo-)N-terminal peptides is essential for identifying protease cleavage sites . We here present an update of a well-established and efficient selection method for enriching N-terminal peptides out of peptide mixtures: N-terminal COFRADIC (COmbined FRActional DIagonal Chromatography). This method is based on the old concept of diagonal chromatography, which involves a peptide modification step in between otherwise identical chromatographic separations, with this modification step finally allowing for the isolation of N-terminal peptides by longer retention of non-N-terminal peptides on the resin. N-terminal COFRADIC has been successfully applied in many protease-centric studies, as well as for studies on protein alpha-N-acetylation and on characterizing alternative translation initiation events.
VIB Proteomics Core, B-9000, Ghent, Belgium.|VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
An
A
Staes
VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
Petra
P
Van Damme
VIB Proteomics Core, B-9000, Ghent, Belgium.|VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
Evy
E
Timmerman
VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
Bart
B
Ruttens
VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
Elisabeth
E
Stes
VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium.
Kris
K
Gevaert
VIB Proteomics Core, B-9000, Ghent, Belgium. francis.impens@vib-ugent.be.|VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium. francis.impens@vib-ugent.be.|Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium. francis.impens@vib-ugent.be.
Francis
F
Impens
D002853:Chromatography, Liquid / Q000379:methods; D010450:Endopeptidases / Q000378:metabolism*; D007553:Isotope Labeling; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D059748:Proteolysis; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012984:Software; D013223:Statistics as Topic / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D057188:Workflow
D010446:Peptide Fragments; D020543:Proteome; D010450:Endopeptidases
Degradomics; N-terminomics; neo-N-termini