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Protease Substrate Profiling by N-Terminal COFRADIC

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Protein Terminal Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1574))

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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Author information

Authors and Affiliations

  1. VIB Proteomics Core, B-9000, Ghent, Belgium

    An Staes, Evy Timmerman & Francis Impens

  2. VIB-UGent Center for Medical Biotechnology, B-9000, Ghent, Belgium

    An Staes, Petra Van Damme, Evy Timmerman, Bart Ruttens, Elisabeth Stes, Kris Gevaert & Francis Impens

  3. Department of Biochemistry, Ghent University, B-9000, Ghent, Belgium

    An Staes, Petra Van Damme, Evy Timmerman, Bart Ruttens, Elisabeth Stes, Kris Gevaert & Francis Impens

Authors
  1. An Staes
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  2. Petra Van Damme
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  3. Evy Timmerman
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  4. Bart Ruttens
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  5. Elisabeth Stes
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  6. Kris Gevaert
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  7. Francis Impens
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Correspondence to Francis Impens .

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Editors and Affiliations

  1. Institute of Molecular Medicine and Cell Research, University Freiburg, Freiburg, Germany

    Oliver Schilling

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Staes, A. et al. (2017). Protease Substrate Profiling by N-Terminal COFRADIC. In: Schilling, O. (eds) Protein Terminal Profiling. Methods in Molecular Biology, vol 1574. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6850-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6850-3_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6849-7

  • Online ISBN: 978-1-4939-6850-3

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