title: Design of selective substrates of proteinase 3 using combinatorial chemistry methods.
abstract: In this study, chemical synthesis of the selective chromogenic/fluorogenic substrates for proteinase 3 is described. The substrates' sequence was obtained using combinatorial chemistry methods. Deconvolution of the tripeptide library against proteinase 3 with general formula ABZ-X3-X2-X1-ANB-NH2 yielded the active sequence. Selected peptide was further modified on its C terminus to investigate the impact of chromophore moiety modification on enzyme-substrate interaction. To determine specificity, activity of selected substrates was characterized against proteinase 3 and neutrophil elastase. Finally, the peptide ABZ-Tyr-Tyr-Abu-ANB-NH2 displayed the highest value of specificity constant (k(cat)/K(M)=189 x 10(3) M(-1) s(-1)) for proteinase 3. To the best of our knowledge, this is the first short peptide that undergoes selective proteolysis by proteinase 3 and displays no significant hydrolysis in the presence of human neutrophil elastase and cathepsin G.
authors.affiliation: Faculty of Chemistry, University of Gdańsk, 80-952 Gdańsk, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wysocka
authors.forename: Adam
authors.initials: A
authors.lastname: Lesner
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Guzow
authors.forename: Lucyna
authors.initials: L
authors.lastname: Mackiewicz
authors.forename: Anna
authors.initials: A
authors.lastname: Legowska
authors.forename: Wiesław
authors.initials: W
authors.lastname: Wiczk
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Rolka
mesh_terms: D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D006801:Humans; D006868:Hydrolysis; D019272:Leukocyte Elastase / Q000378:metabolism; D013058:Mass Spectrometry; D053146:Myeloblastin / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D017186:Titrimetry
chemical_list: D010455:Peptides; D019272:Leukocyte Elastase; D053146:Myeloblastin