Purification and characterization of myofibril-bound serine proteinase from carp Cyprinus carpio ordinary muscle

Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
K OsatomiT Ishihara

Abstract

1. A novel myofibril-bound serine proteinase (MBP) has been purified from ordinary muscle of the carp Cyprinus carpio. 2. It was solubilized from the myofibril fraction with acid treatment (under the conditions of 0.6 M KCl, pH 4.0), then purified by column chromatographic steps on Ultrogel AcA 54, and Arginine-Sepharose 4B. 3. The purified enzyme revealed a single protein band on SDS-PAGE, and its molecular mass was estimated to be 30 kDa by SDS-PAGE and gel filtration. 4. The optimum pH and temperature of the enzyme were 8.0 and 55 degrees C, respectively, when Boc-Phe-Ser-Arg-MCA and casein were used as substrates. 5. The enzyme hydrolyzed Boc-Gln-Arg-Arg-MCA most rapidly, and also hydrolyzed the substrates for trypsin-type proteinase, but not for chymotrypsin. The enzyme was inhibited by serine proteinase inhibitors such as DFP, STI and leupeptin. These results suggested that the enzyme was a trypsin-type serine proteinase. 6. Boc-Phe-Ser-Arg-MCA hydrolyzing activity of the purified enzyme was reduced by addition of NaCl, but the caseinolytic activity and Boc-Phe-Ser-Arg-MCA hydrolyzing activity of the partially purified enzyme were activated by NaCl.

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Citations

Dec 31, 2003·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboTadashi Ishihara
Dec 3, 1999·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·M J CaoT Ishihara
May 19, 2000·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·M J CaoT Ishihara
Feb 13, 2001·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·M J CaoT Ishihara
Mar 30, 2002·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Marcela P SangorrínJorge J Sánchez
Apr 3, 2003·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Soottawat BenjakulKittima Leelapongwattana
Jan 4, 2001·The International Journal of Biochemistry & Cell Biology·M P SangorrínJ J Sánchez
Jun 8, 2001·The International Journal of Biochemistry & Cell Biology·M P SangorrínJ J Sánchez
Jun 2, 2000·Biochemical and Biophysical Research Communications·M J CaoT Ishihara
Jan 15, 2013·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Cui-Hong DuMin-Jie Cao
Apr 26, 2011·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Qiu-Feng CaiMin-Jie Cao
Dec 1, 2009·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Guo-Ping WuMin-Jie Cao
Jun 30, 2009·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Shonisani C TshidinoRyno J Naudé
Nov 17, 2004·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Thomas P Mommsen
Dec 8, 2004·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboTadashi Ishihara
Jun 9, 2005·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboFutoshi Aranishi
Dec 31, 2016·Food Science and Biotechnology·Chen-Chen ZhaoYue Cha
Oct 29, 2020·Journal of Agricultural and Food Chemistry·Yuri KominamiHideki Ushio

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