Reddy S R, Watts D C
Department of Biochemistry, Guy's Hospital Medical School, London, U.K.
Comp Biochem Physiol Biochem Mol Biol. 1994 May;108(1):73-8. doi: 10.1016/0305-0491(94)90167-8.
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.
二聚体兔肌肌酸激酶（MM-CK）通过其一个亚基（MM-CKA）与溴化氰活化的琼脂糖4B结合。用盐酸胍处理MM-CKA可释放未结合的亚基，从而产生基质结合的单体（M-CKB）。M-CKB与解离的MM-CK可溶性亚基重新组合，以重构基质结合的二聚体（MM-CKC）。M-CKB还与兔脑粗提物中的BB-CK解离亚基以及海参肌肉中的精氨酸激酶（MM-AK）结合，分别形成基质结合的杂合二聚体MB-CKC和M-CK/M-AKC。盐酸胍梯度洗脱研究表明MM-CKA、MM-CKC和MB-CKC在相同浓度的变性剂（0.96 M）下均会解离，而M-CK/M-AKC杂合二聚体稳定性较差，在0.5 M时解离。棘皮动物和哺乳动物磷酸原激酶亚基之间形成具有双重底物特异性的杂合酶的特异性相互作用支持了这些酶具有共同进化起源的观点。
