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

  • Marcin Poręba
  • Aleksandra Szalek
  • Paulina Kasperkiewicz
  • Marcin Drąg
Part of the Methods in Molecular Biology book series (MIMB, volume 1133)


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.

Key words

Substrate specificity Caspase Fluorogenic substrate Combinatorial library Cysteine protease 



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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Marcin Poręba
    • 1
  • Aleksandra Szalek
    • 1
  • Paulina Kasperkiewicz
    • 1
  • Marcin Drąg
    • 1
  1. 1.Division of Bioorganic Chemistry, Faculty of ChemistryWroclaw University of TechnologyWroclawPoland

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