caa a22 4500 397557450 CHVBK 20180308164806.0 cr unu---uuuuu 161202e199604 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021289 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021289 Purification and Characterization of Novel Trypsin-Like Serine Proteases from Mouse Spleen [Elektronische Daten] [Naomi Fukusen, Yosuke Aoki] Novel trypsin-like serine proteases (mouse trypsin-type serine proteases 1 and 2 [MTSP-1 and -2]) were purified to homogeneity from mouse spleen. Each protease consisted of a single polypeptide with a molecular mass of about 29 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions. Both were totally inhibited by diisopropylfluorophosphate, soybean trypsin inhibitor, aprotinin, antipain, and leupeptin and partially inhibited by chymostatin and dithiothreitol, suggesting that they are trypsin-like serine proteases. They hydrolyzed synthetic substrates for trypsin-like proteases but not those for chymotrypsin-like proteases, elastase and kalli-krein. MTSP-1 hydrolyzed terf-butyloxycarbonyl (Boc)-Asp(OBzl)Pro-Arg-amino-4-methyl-coumaryl-7-amide (MCA) and Boc-De-Glu-Gly-Arg-MCA faster than Boc-Phe-Ser-Arg-MCA. On the other hand, MTSP-2 hydrolyzed Boc-Phe-Ser-Arg-MCA most rapidly, with a specific activity 15 times higher than that of MTSP-1. The N-terminal amino acid sequence of MTSP-1 was Ile-Val-Gly-Gly-Tyr-Thr-His-Leu-Asp-Asn-Gln-Val-Pro-Tyr. This sequence was 71% homologous with the N-terminal of bovine trypsin. The Boc-Phe-Ser-Arg-MCA hydrolyzing activity of mouse spleen significantly (p < 0.01) increased to about 1.5-fold the basal activity 2 weeks after an injection of Freund's complete adjuvant, suggesting that these proteases are involved in the immune response. © by the Journal of Biochemistry Regular Papers nationallicence adjuvant nationallicence immune response nationallicence mouse spleen nationallicence serine protease nationallicence trypsin-like nationallicence Fukusen Naomi Department of Biochemistry and Nutrition, National Institute of Public Health, 6-1 Shirokanedai 4-chome, Minato-ku, Tokyo 108 aut Aoki Yosuke Department of Biochemistry and Nutrition, National Institute of Public Health, 6-1 Shirokanedai 4-chome, Minato-ku, Tokyo 108 aut The Journal of Biochemistry Oxford University Press 119/4(1996-04), 633-638 0021-924X 119:4<633 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021289 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021289 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fukusen Naomi Department of Biochemistry and Nutrition, National Institute of Public Health, 6-1 Shirokanedai 4-chome, Minato-ku, Tokyo 108 aut NATIONALLICENCE 700 1- Aoki Yosuke Department of Biochemistry and Nutrition, National Institute of Public Health, 6-1 Shirokanedai 4-chome, Minato-ku, Tokyo 108 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/4(1996-04), 633-638 0021-924X 119:4<633 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford