The Simultaneous Determination of Intracellular pH and Cell Energy Status

Abstract

The relationship between the apparent equilibrium constant of creatine kinase and intracellular pH was evaluated in CHO and murine FSaII tumor cells. The apparent equilibrium constant, K' = [ATP][Cr]/[ADP][PCr], was determined from acid extracts at variable pH. Intracellular pH (pHi) was determined from the intracellular/extracellular distribution of the weak acid 5,5-dimethyl-2,4-oxazolidinedione. Over the intracellular pH range of 7.2 to 6.1, K' increased by a factor of approximately 10. Intracellular pH was related to the apparent equilibrium constant by the equation pHi = -log K' + log K, where the value of the constant log (log[K'/H+]) was 8.09. Over the same pH range, the concentration of phosphocreatine decreased with pH. Essentially identical results were obtained in CHO and FSaII tumor cells. The similar apparent equilibrium constants in CHO and FSaII cells suggest that assessment of the creatine kinase metabolites will be useful not only for determination of cell energy status but also for the determination of intracellular pH. This information may be useful for the design of therapeutic strategies which are influenced by pH or energy status such as hyperthermia, and drugs which are weak acids or bases, including hypoxic cell radiosensitizers.