Enzyme termini of a phosphocreatine shuttle. Purification and characterization of two creatine kinase isozymes from sea urchin sperm.

摘要: Two isozymes of creatine kinase have been purified from sperm the sea urchin, Strongylocentrotus purpuratus. One isozyme was flagellum, and other head. Both require nonionic detergent for extraction sperm. The flagellar is a monomeric species with an Mr 145,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 126,000 sucrose density gradient filtration analyses. Creatine heads localized to mitochondrion antibody raised against mouse muscle kinase. This mitochondrial multimeric, 47,000 electrophoresis, but 240,000 native enzyme. Peptide mapping indicates that two are not related. following kinetic characteristics were observed isozymes, respectively. In direction ATP formation, at pH 6.6 25 degrees C, specific activities 235 180 units/mg; optima 6.7 6.9 Michaelis constants 0.13 0.055 mM ADP 5.8 2.7 phosphocreatine. phosphocreatine 7.5 29 47 7.7 0.89 0.31 39 62 creatine. These unique constitute termini shuttle urchin responsible energy transport distal flagellum (Tombes, R. M., Shapiro, B. M. (1985) Cell 41, 325-334; Tombes, Brokaw, C. J., (1987) Biophys. 52, 75-86).