The temperature dependence of intracellular pH in isolated frog skeletal muscle: lessons concerning the Na(+)-H+ exchanger.
We used 31P NMR to investigate the temperature-dependence of intracellular pH (pHi) in isolated frog skeletal muscles. We found that Ln[H+i] is a linear function of 1/Tabs paralleling those of neutral water (i.e., H+ = OH-) and of a solution containing the fixed pH buffers of frog muscle cytosol. This classical van't Hoff relationship was unaffected by inhibition of glycolysis and was not dependent upon the pH or [Na+] in the bathing solution. Insulin stimulation of Na(+)-H+ exchange shifted the intercept in the alkaline direction but had not effect on the slope. Acid loading followed by washout resulted in an amiloride-sensitive return to the (temperature dependent) basal pHi. These results show that the temperature dependence of activation of Na(+)-H+ exchange is similar to that of the intracellular buffers, and suggest that constancy of [H+]/ [OH-] with changing temperature is achieved in the short term by intracellular buffering and in the long term by the set-point of the Na(+)-H+ exchanger. Proton activation of the exchanger has an apparent standard enthalpy change (delta H degree) under both control and insulin-stimulated conditions that is similar to the delta H degree of the intracellular buffers and approximately half of the delta H degree for the dissociation of water. Thus, the temperature-dependent component of the standard free-energy change (delta F degree) is unaffected by insulin stimulation, suggesting that changes in Arrhenius activation energy (Ea) may not be a part of the mechanism of hormone stimulation.
Marjanovic
M
M
Department of Zoology, Eastern Illinois University, Charleston 61920, USA.
Elliott
A C
AC
Dawson
M J
MJ
D000143:Acids / Q000378:metabolism; D000818:Animals; D002138:Calibration; D006863:Hydrogen-Ion Concentration; D007424:Intracellular Fluid; D018482:Muscle, Skeletal / Q000378:metabolism*; D011522:Protons; D011894:Rana pipiens; D011896:Rana temporaria; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013696:Temperature
D000143:Acids; D011522:Protons; D017923:Sodium-Hydrogen Exchangers