Hybridization of matrix-bound MM-creatine kinase with BB-creatine kinase and arginine kinase.
Dimeric rabbit muscle creatine kinase (MM-CK) was bound to CNBr-activated Sepharose 4B by one of its subunits (MM-CKA). Treatment of MM-CKA with guanidine hydrochloride released the unbound subunit to yield the matrix-bound monomer (M-CKB). M-CKB recombined with dissociated MM-CK soluble subunits to reconstitute a matrix-bound dimer (MM-CKC). M-CKB also associated with dissociated subunits of BB-CK from crude extracts of rabbit brain and of arginine kinase from sea cucumber muscle (MM-AK) to form the matrix-bound heterohybrids MB-CKC and M-CK/M-AKC, respectively. Guanidine hydrochloride gradient elution studies showed that MM-CKA, MM-CKC and MB-CKC were all dissociated at the same concentration of the denaturant (0.96 M), while the M-CK/M-AKC heterohybrid was less stable, dissociating at 0.5 M. The specific interaction between subunits of echinoderm and mammalian phosphagen kinases to form a hybrid enzyme of dual substrate specificity supports the view that these enzymes had a common evolutionary origin.
Reddy
S R
SR
Department of Biochemistry, Guy's Hospital Medical School, London, U.K.
Watts
D C
DC
D000818:Animals; D001122:Arginine Kinase / Q000737:chemistry* / Q000302:isolation & purification; D005075:Biological Evolution; D001921:Brain / Q000201:enzymology; D003402:Creatine Kinase / Q000737:chemistry* / Q000302:isolation & purification; D007527:Isoenzymes; D009132:Muscles / Q000201:enzymology; D011487:Protein Conformation; D011817:Rabbits; D012616:Sea Cucumbers / Q000201:enzymology; D013045:Species Specificity
D007527:Isoenzymes; D003402:Creatine Kinase; D001122:Arginine Kinase