title: In silico design, synthesis, and assays of specific substrates for proteinase 3: influence of fluorogenic and charged groups.
abstract: Neutrophil serine proteases are specific regulators of the immune response, and proteinase 3 is a major target antigen in antineutrophil cytoplasmic antibody-associated vasculitis. FRET peptides containing 2-aminobenzoic acid (Abz) and N-(2,4-dinitrophenyl)ethylenediamine (EDDnp) as fluorophore and quencher groups, respectively, have been widely used to probe proteases specificity. Using in silico design followed by enzymatic assays, we show that Abz and EDDnp significantly contribute to substrate hydrolysis by PR3. We also propose a new substrate specific for PR3.
authors.affiliation: Department of Molecular Biology, University of Bergen , 5008, Bergen, Norway.
authors.forename: Shailesh
authors.initials: S
authors.lastname: Narawane
authors.forename: Adnan
authors.initials: A
authors.lastname: Budnjo
authors.forename: Cédric
authors.initials: C
authors.lastname: Grauffel
authors.forename: Bengt Erik
authors.initials: BE
authors.lastname: Haug
authors.forename: Nathalie
authors.initials: N
authors.lastname: Reuter
mesh_terms: D003198:Computer Simulation; D015195:Drug Design; D057075:Enzyme Assays; D005029:Ethylenediamines / Q000138:chemical synthesis* / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D006868:Hydrolysis; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D053146:Myeloblastin / Q000737:chemistry*; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D062367:ortho-Aminobenzoates / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: D005029:Ethylenediamines; D009842:Oligopeptides; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; C026951:N-(2,4-dinitrophenyl)ethylenediamine; D053146:Myeloblastin