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.
CITATION STYLE
Staes, A., van Damme, P., Timmerman, E., Ruttens, B., Stes, E., Gevaert, K., & Impens, F. (2017). Protease substrate profiling by N-terminal COFRADIC. In Methods in Molecular Biology (Vol. 1574, pp. 51–76). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6850-3_5
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