Department of Molecular Biology, University of Bergen , 5008, Bergen, Norway.
J Med Chem. 2014 Feb 13;57(3):1111-5. doi: 10.1021/jm401559r. Epub 2014 Jan 21.
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.
中性粒细胞丝氨酸蛋白酶是免疫反应的特异性调节剂，蛋白酶 3 是抗中性粒细胞胞质抗体相关性血管炎的主要靶抗原。含有 2-氨基苯甲酸 (Abz) 和 N-(2,4-二硝基苯)乙二胺 (EDDnp) 作为荧光团和猝灭基团的 FRET 肽已被广泛用于探测蛋白酶的特异性。通过计算机设计和酶促实验，我们表明 Abz 和 EDDnp 显著促进 PR3 的底物水解。我们还提出了一种新的 PR3 特异性底物。
