Purification and partial characterization of a myofibril-bound serine protease from ostrich skeletal muscle

A myofibril-bound serine protease (MBSP) was partially purified from ostrich (Struthio camelus) skeletal muscle. MBSP was dissociated from the myofibrillar fraction by ethylene glycol treatment at pH 8.5, followed by partial purification via Toyopearl Super Q 650S and p-aminobenzamidine column chromatographies. Ostrich MBSP revealed a major protein band of approximately 21kDa on SDS-PAGE, showing proteolytic activity after casein zymography. Optima pH and temperature of ostrich MBSP were 8 and 40[degrees]C, respectively. Substrate specificity analysis revealed that the enzyme cleaved synthetic fluorogenic substrates at the carboxyl side of arginine residues. Kinetic parameters (K.sub.m and V.sub.max values) were calculated from Lineweaver-Burk plots. The kinetic characteristics of ostrich MBSP were compared to values obtained for commercial bovine trypsin in this study, as well as those obtained for MBSP from mouse and various fish species. The results suggest that ostrich MBSP is a tryptic-like serine protease. Ostrich MBSP exhibited low sequence identity to commercial bovine trypsin (44%), MBSP from lizard fish skeletal muscle (33%) and trypsinogen from ostrich pancreas (22%).

Author Affiliation:

(a) Department of Biochemistry and Microbiology, Nelson Mandela Metropolitan University, P.O. Box 77000, Summerstrand South campus, Port Elizabeth, 6031, South Africa

(b) Graduate School of Life Sciences, Tohoku University, Laboratory of Biomolecular Function, Sendai, 981-8555, Japan

Article History:

Received 1 April 2009; Revised 13 June 2009; Accepted 14 June 2009

Source Citation:Tshidino, Shonisani C., Jason Krause, Abayomi P. Adebiyi, Koji Muramoto, and Ryno J. Naude. "Purification and partial characterization of a myofibril-bound serine protease from ostrich skeletal muscle.(Report)." Comparative Biochemistry and Physiology, Part B 154.2 (Oct 2009): 229(6). Academic OneFile. Gale. BROWARD COUNTY LIBRARY. 3 Aug. 2009
.
Gale Document Number:A204426537

Disclaimer:This information is not a tool for self-diagnosis or a substitute for professional care.