title: Substrate specificity determination of mouse implantation serine proteinase and human kallikrein-related peptidase 6 by phage display.
abstract: We constructed a random library of hexapeptides displayed on the surface of bacteriophage T7 to determine the substrate specificity of proteinases. The phage-displayed library was subjected to repeated rounds of biopanning with native implantation serine proteinase and recombinant human kallikrein-related peptidase 6 (KLK6) followed by selection and identification of putative substrates. For both enzymes, the results obtained demonstrate a preference for arginine and lysine at multiple positions in the recognition cleavage motif, confirming their previously reported trypsin-like substrate specificity. In the case of KLK6, there is also a pronounced presence of tryptophan within the cleaved peptide sequences, indicating its potential dual substrate specificity, acting as both a trypsin and chymotrypsin-like enzyme.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Calgary, Calgary T2N 4N1, Canada.
authors.forename: Navneet
authors.initials: N
authors.lastname: Sharma
authors.forename: Katerina
authors.initials: K
authors.lastname: Oikonomopoulou
authors.forename: Kenichi
authors.initials: K
authors.lastname: Ito
authors.forename: Bernard
authors.initials: B
authors.lastname: Renaux
authors.forename: Eleftherios P
authors.initials: EP
authors.lastname: Diamandis
authors.forename: Morley D
authors.initials: MD
authors.lastname: Hollenberg
authors.forename: Derrick E
authors.initials: DE
authors.lastname: Rancourt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D051379:Mice; D019151:Peptide Library; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D017186:Titrimetry
chemical_list: D019151:Peptide Library; D012697:Serine Endopeptidases