Interactions of temperature and pH on the regulatory properties of pyruvate kinase from organs of a marine mollusc
The effects of low temperature assay (5 °C) on the properties of the aerobic (low phosphate) vs. anoxic (high phosphate) forms of pyruvate kinase (PK) from foot muscle and gill of the whelk Busycon canaliculatum (L.) were assessed at two pH values, pH 7.00 and 7.25, and compared to control conditions of 20 °C and pH 7.00 (all assayed in imidazole buffer). When pH was held constant at 7.00, the decrease in assay temperature to 5 °C had large effects on the measured kinetic parameters of all PK forms, as compared to 20 °C and pH 7.00. However, when assay pH was allowed to rise, from 7.00 to 7.25, with the temperature decrease to 5 °C there were fewer alterations of kinetic parameters and quantitatively smaller changes to enzyme properties. It appears, then, that when pH rises with decreasing temperature following alphastat predictions, kinetic properties of PK are largely conserved. Low temperature, at either pH value, had several significant effects on PK properties. For example, low temperature raised the S0.5 for phosphoenolpyruvate of PK-anoxic from gill by 3-6 fold and decreased the I50 Mg · ATP for PK-anoxic from foot by the same amount. Arrhenius plots of PK activity for the gill PK forms showed a distinct break at 10 °C; > 10 °C Q10 was 2.5 whereas < 10 °C Q10 was 8.4. Temperature-dependent changes in all cases affected enzyme properties in a manner that would restrict enzyme function at low temperature.
|Keywords||Busycon canaliculatum, pH effects on pyruvate kinase, Pyruvate kinase, Temperature effects on pyruvate kinase|
|Journal||Journal of Experimental Marine Biology and Ecology|
Michaelidis, B. (Basile), & Storey, K. (1990). Interactions of temperature and pH on the regulatory properties of pyruvate kinase from organs of a marine mollusc. Journal of Experimental Marine Biology and Ecology, 140(3), 187–196. doi:10.1016/0022-0981(90)90126-W