Isoform specific interactions of troponin I and troponin C determine pH sensitivity of myofibrillar Ca2+ activation.
We investigated whether differences in isoforms of troponin I (TnI) and troponin C (TnC) can account for the greater inhibition of Ca(2+)-dependent MgATPase activity by acidic pH in cardiac (c) than in fast skeletal (fs) myofilaments. We studied fast skeletal myofibrils from which whole Tn was extracted by displacement with excess fsTnT (the tropomyosin binding subunit of Tn) followed by reconstitution with TnC-TnI. Exchange of fsTnI with cTnI did not alter the effect of a drop in pH from 7.0 to 6.5 on the relation between pCa (-log[Ca2+]) and MgATPase activity of fast skeletal myofibrils. Exchange of fsTnC with cTnC did, however, induce an increase in the effect of this same pH change on Ca2+ activation. Yet, the pH sensitivity of Ca2+ activation of fast skeletal myofibrils containing cTnC was not as great as that of native cardiac myofibrils. However, when both fsTnC and fsTnI of fast skeletal myofibrils were replaced by cTnC-cTnI, there was a pH-induced shift in Ca2+ sensitivity similar to that of cardiac myofibrils. In studies using fluorescent probes, both pure fsTnC and pure cTnC showed decreased Ca2+ binding as pH was lowered. This decrease was potentiated in the fsTnC-fsTnI and cTnC-cTnI complexes. However, the effect of acidic pH was the same in fsTnC and the hybrid complex, fsTnC-cTnI, and in cTnC and the hybrid complex, cTnC-fsTnI. Thus, isoform specific interactions between TnI and TnC appear important in the differential response of skeletal and cardiac myofilaments to acidosis.
Department of Physiology and Biophysics, University of Illinois College of Medicine, Chicago 60612-7342.
K L
KL
Ball
M D
MD
Johnson
R J
RJ
Solaro
D000818:Animals; D001665:Binding Sites; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D004789:Enzyme Activation; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009210:Myofibrils / Q000201:enzymology*; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D014336:Troponin / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D019209:Troponin C; D019210:Troponin I
D005456:Fluorescent Dyes; D014336:Troponin; D019209:Troponin C; D019210:Troponin I; D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium