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Bacterial Cell Wall Homeostasis pp 99–108Cite as

Liquid Chromatography-Tandem Mass Spectrometry to Define Sortase Cleavage Products

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1440))

Abstract

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.

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Acknowledgements

This work was supported by a CIHR grant (MOP—137004).

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    Authors and Affiliations

    1. Department of Biology, McMaster University, Hamilton, ON, Canada

      Andrew Duong, Danielle L. Sexton & Marie A. Elliot

    2. Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada

      Andrew Duong, Kalinka Koteva & Danielle L. Sexton

    3. Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada

      Kalinka Koteva

    4. Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Life Science Building, RM 329, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada

      Marie A. Elliot

    Authors
    1. Andrew Duong
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    2. Kalinka Koteva
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    3. Danielle L. Sexton
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    4. Marie A. Elliot
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    Corresponding author

    Correspondence to Marie A. Elliot .

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    Editors and Affiliations

    1. Dept. of Biochem & Dept. of Bio and Med., Univ. of Cambridge andOxford Brookes Unv, Cambridge, United Kingdom

      Hee-Jeon Hong

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    Duong, A., Koteva, K., Sexton, D.L., Elliot, M.A. (2016). Liquid Chromatography-Tandem Mass Spectrometry to Define Sortase Cleavage Products. In: Hong, HJ. (eds) Bacterial Cell Wall Homeostasis. Methods in Molecular Biology, vol 1440. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3676-2_8

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    • DOI: https://doi.org/10.1007/978-1-4939-3676-2_8

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-3674-8

    • Online ISBN: 978-1-4939-3676-2

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