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Positional Scanning Substrate Combinatorial Library (PS-SCL) Approach to Define Caspase Substrate Specificity

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Caspases,Paracaspases, and Metacaspases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1133))

Abstract

Positional scanning substrate combinatorial library (PS-SCL) is a powerful tool for studying substrate specificity of proteolytic enzymes. Here, we describe the protocol for analyzing S4-S2 pockets preferences of caspases using PS-SCL. Additionally, we describe procedures for the identification of optimal substrates sequence after PS-SCL, solid phase synthesis, and purification of selected fluorogenic substrates, as well as their kinetic analysis.

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Acknowledgments

This work was supported by the National Science Centre grant 2011/03/B/ST5/01048 and the Foundation for Polish Science in Poland. This work is co-financed by the European Union as part of the European Social Fund.

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Authors and Affiliations

  1. Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland

    Marcin Poręba, Aleksandra Szalek, Paulina Kasperkiewicz & Marcin Drąg

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  1. Marcin Poręba
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  2. Aleksandra Szalek
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  3. Paulina Kasperkiewicz
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  4. Marcin Drąg
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Editor information

Editors and Affiliations

  1. Dept of Plant Biology & Forest Genetics, Swedish Univ of Agricultural Sciences Linnean Center for Plant Biology, Uppsala, Sweden

    Peter V. Bozhkov

  2. Sanford-Burnham Medical Research Inst. Apoptosis & Cell Death Research Prgm, La Jolla, California, USA

    Guy Salvesen

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Poręba, M., Szalek, A., Kasperkiewicz, P., Drąg, M. (2014). Positional Scanning Substrate Combinatorial Library (PS-SCL) Approach to Define Caspase Substrate Specificity. In: V. Bozhkov, P., Salvesen, G. (eds) Caspases,Paracaspases, and Metacaspases. Methods in Molecular Biology, vol 1133. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0357-3_2

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  • DOI: https://doi.org/10.1007/978-1-4939-0357-3_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0356-6

  • Online ISBN: 978-1-4939-0357-3

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