| Chapter title |
Protease Substrate Profiling by N-Terminal COFRADIC
|
|---|---|
| Chapter number | 5 |
| Book title |
Protein Terminal Profiling
|
| Published in |
Methods in molecular biology, March 2017
|
| DOI | 10.1007/978-1-4939-6850-3_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6849-7, 978-1-4939-6850-3
|
| Authors |
Staes, An, Van Damme, Petra, Timmerman, Evy, Ruttens, Bart, Stes, Elisabeth, Gevaert, Kris, Impens, Francis, An Staes, Petra Van Damme, Evy Timmerman, Bart Ruttens, Elisabeth Stes, Kris Gevaert, Francis Impens |
| Editors |
Oliver Schilling |
| Abstract |
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. |
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