title: Positional scanning substrate combinatorial library (PS-SCL) approach to define caspase substrate specificity.
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.
authors.affiliation: Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Poręba
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Szalek
authors.forename: Paulina
authors.initials: P
authors.lastname: Kasperkiewicz
authors.forename: Marcin
authors.initials: M
authors.lastname: Drąg
mesh_terms: D000595:Amino Acid Sequence; D020169:Caspases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020650:Combinatorial Chemistry Techniques; D006801:Humans; D007700:Kinetics; D008967:Molecular Biology / Q000379:methods*; D019151:Peptide Library; D013379:Substrate Specificity
chemical_list: D019151:Peptide Library; D020169:Caspases