Ca2+ binding to cardiac troponin C: effects of temperature and pH on mammalian and salmonid isoforms

Abstract

A reduction in temperature lowers the Ca2+ sensitivity of skinned cardiac myofilaments but this effect is attenuated when native cardiac troponin C (cTnC) is replaced with skeletal TnC. This suggests that conformational differences between the two isoforms mediate the influence of temperature on contractility. To investigate this phenomenon, the functional characteristics of bovine cTnC (BcTnC) and that from rainbow trout, Oncorhynchus mykiss, a cold water salmonid (ScTnC), have been compared. Rainbow trout maintain cardiac function at temperatures cardioplegic to mammals. To determine whether ScTnC is more sensitive to Ca2+ than BcTnC, F27W mutants were used to measure changes in fluorescence with in vitro Ca2+ titrations of site II, the activation site. When measured under identical conditions, ScTnC was more sensitive to Ca2+ than BcTnC. At 21°C, pH 7.0, as indicated by K 1/2 (−log[Ca] at half-maximal fluorescence, where [Ca] is calcium concentration), ScTnC was 2.29-fold more sensitive to Ca2+ than BcTnC. When pH was kept constant (7.0) and temperature was lowered from 37.0 to 21.0°C and then to 7.0°C, the K 1/2 of BcTnC decreased by 0.13 and 0.32, respectively, whereas the K 1/2 of ScTnC decreased by 0.76 and 0.42, respectively. Increasing pH from 7.0 to 7.3 at 21.0°C increased the K 1/2 of both BcTnC and ScTnC by 0.14, whereas the K 1/2 of both isoforms was increased by 1.35 when pH was raised from 7.0 to 7.6 at 7.0°C.