Transient-Phase Studies on the Creatine Kinase Reaction. The Analysis of a Reaction Pathway with Three Intermediates

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Abstract
The initial formation of creatine phosphate by [rabbit muscle] creatine kinase was studied in the ms range and the effect of temperature on the transient and steady-state phases exploited. At 25 and 35.degree. C there was no transient phase. At 4.degree. C the time course of creatine phosphate formation was complex and consisted of 3 transient phases: a lag phase, a burst phase and a steady-state phase. A reaction scheme for creatine kinase which includes 3 intermediates was proposed. Despite the completeness of the time course, the extraction of estimates for the rate constants was difficult and computer simulation and iterative methods were used. Attempts were made to provide evidence for the complex enzyme.cntdot.ADP.cntdot.metaphosphate creatine on the creatine kinase reaction pathway. Under the conditions used these attempts were unsuccessful at times down to 2.5 ms, at 4 or 35.degree. C.

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