Isolation of three creatine kinases MM from porcine skeletal muscle.
The purified creatine kinase MM of porcine skeletal muscle [Takasawa, T. & Shiokawa, H. (1981) J. Biochem. 90, 195-204] was separated into three distinct fractions by isoelectric focusing (IEF) in a sucrose gradient column, and the three active fractions were isolated by repeated IEF. There were one major fraction with isoelectric point (pI) 6.57 and two minor fractions with pI 6.74 and pI 6.34, respectively. No differences were observed in the IEF pattern of the enzyme in the presence and absence of dithiothreitol throughout the column. There was no interconversion from one form to another during IEF. The distribution of the three forms on IEF was not affected by adding protease inhibitor to the extraction medium. Of the three fractions, the major fraction had the highest specific activity. The three fractions differed from one another in their amino acid compositions. Not only porcine muscle but also rabbit muscle creatine kinase displayed this type of heterogeneity. Such microheterogeneities may occur widely in muscle creatine kinases.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003402:Creatine Kinase / Q000302:isolation & purification* / Q000378:metabolism; D007527:Isoenzymes; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D013552:Swine
D000596:Amino Acids; D007527:Isoenzymes; D003402:Creatine Kinase