Comparison of kinetic constants of creatine kinase isoforms.

The purpose of this study was to elucidate the functional differences between the CK isoforms by cloning the cDNAs of 12 CK isoforms: the M and B cytoplasmic forms and uMiCK from mouse, the M1, M2 and B cytoplasmic forms from Danio rerio, M1 and M2 cytoplasmic forms from the lower vertebrate Lampetra japonica, a cytoplasmic CK and a MiCK from the marine worm Neanthes diversicolor, and a cytoplasmic CK and a MiCK from the soft coral Dendronephthya gigantea. These were expressed in Escherichia coli as a fusion protein with maltose-binding protein, and kinetic constants (K(m), K(d) and k(cat)) of all the recombinant enzymes, except for the unstable Dendronephthya cytoplasmic CK, were determined for the forward reaction. The kinetic constants of the M- and B-forms of the mouse and Danio cytoplasmic CKs differed significantly, with the K(m) for creatine (K(m)Cr) of M-CK being three- to nine-fold higher than that of B-CK, possibly reflecting differences in the concentration of creatine in muscle and brain cells.