title: Selection of new chromogenic substrates of serine proteinases using combinatorial chemistry methods.
abstract: Chemical synthesis, physicochemical characterization and kinetic investigations of a tetrapeptide library of chromogenic substrates containing the amide of 5-amino-2-nitrobenzoic acid (Anb(5,2)-NH(2)) at their C-termini are reported. Anb(5,2)-NH(2) served as a chromophore released upon enzymatic action. The library consisting of 9567 peptides was synthesized using the portioning-mixing method and was screened against bovine a-chymotrypsin and human leukocyte elastase in solution applying an iterative approach. The selected chromogenic substrates were resynthesized and further modified at their N- and C-termini. Finally, two sequences, Z-Phe-Ala-Thr-Tyr-Anb(5,2)-NH(2) and Z-Phe-Phe-Pro-Val-Anb(5,2)-NH(2), were obtained as highly specific substrates for bovine alpha-chymotrypsin and human leukocyte elastase, respectively. The method of synthesis and selection of chromogenic substrates of serine proteinases described herein is straightforward and can be applied to design substrates for other proteases.
authors.affiliation: Bioorganic Chemistry Department, Faculty of Chemistry, University of Gdańsk, ul. Sobieskiego 18, 80-952 Gdańsk, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wysocka
authors.forename: Bozena
authors.initials: B
authors.lastname: Kwiatkowska
authors.forename: Marek
authors.initials: M
authors.lastname: Rzadkiewicz
authors.forename: Adam
authors.initials: A
authors.lastname: Lesner
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Rolka
mesh_terms: D000818:Animals; D002417:Cattle; D002863:Chromogenic Compounds / Q000138:chemical synthesis / Q000378:metabolism*; D002918:Chymotrypsin / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis; D019151:Peptide Library; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002863:Chromogenic Compounds; D009842:Oligopeptides; D019151:Peptide Library; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D019272:Leukocyte Elastase