TY - JOUR
T1 - Purification and partial characterization of a myofibril-bound serine protease from ostrich skeletal muscle
AU - Tshidino, Shonisani C.
AU - Krause, Jason
AU - Adebiyi, Abayomi P.
AU - Muramoto, Koji
AU - Naudé, Ryno J.
N1 - Funding Information:
The authors gratefully acknowledge the financial support from the National Research Foundation (South Africa) and the Japanese Society for the Promotion of Science (Japan) and express their sincere gratitude to the Grahamstown ostrich abattoir (South Africa), for their generous supply of ostrich skeletal muscle.
PY - 2009/10
Y1 - 2009/10
N2 - 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 650 S and p-aminobenzamidine column chromatographies. Ostrich MBSP revealed a major protein band of approximately 21 kDa on SDS-PAGE, showing proteolytic activity after casein zymography. Optima pH and temperature of ostrich MBSP were 8 and 40 °C, respectively. Substrate specificity analysis revealed that the enzyme cleaved synthetic fluorogenic substrates at the carboxyl side of arginine residues. Kinetic parameters (Km and Vmax 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%).
AB - 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 650 S and p-aminobenzamidine column chromatographies. Ostrich MBSP revealed a major protein band of approximately 21 kDa on SDS-PAGE, showing proteolytic activity after casein zymography. Optima pH and temperature of ostrich MBSP were 8 and 40 °C, respectively. Substrate specificity analysis revealed that the enzyme cleaved synthetic fluorogenic substrates at the carboxyl side of arginine residues. Kinetic parameters (Km and Vmax 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%).
KW - Bovine trypsin
KW - Myofibril-bound serine protease
KW - Myofibrils
KW - Ostrich
UR - http://www.scopus.com/inward/record.url?scp=67651125127&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67651125127&partnerID=8YFLogxK
U2 - 10.1016/j.cbpb.2009.06.007
DO - 10.1016/j.cbpb.2009.06.007
M3 - Article
C2 - 19559097
AN - SCOPUS:67651125127
SN - 1096-4959
VL - 154
SP - 229
EP - 234
JO - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
JF - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
IS - 2
ER -