[The mechanism of thermo-induced functional uncoupling of the Ca-pump in the sarcoplasmic reticulum of skeletal muscles].
Short-term heating at 45 degrees C of diluted suspensions of rabbit skeletal muscle sarcoplasmic reticulum (SR) vesicles sharply reduces the Ca/ATP ratio without causing any appreciable changes in the Ca-ATPase activity. The mechanism underlying the uncoupling effect of thermoinactivation was investigated. It was shown that the decrease in the effectiveness of Ca2+ accumulation is correlated with the level of Ca2+ accumulated by SR vesicles. At the same time, thermal inactivation has no effect on passive permeability of SR membranes for Ca2+. Ca2+ (100 microM) added to the inactivation medium partly protects the enzyme against uncoupling. Thermal inactivation of the SR Ca-pump is concomitant with a decreased microviscosity of SR membranes as measured by pyrene excimerization. Addition of 100 microM Ca2+ to the thermoinactivation medium prevents the decrement of microviscosity. It was found that N-3-pyrenylmaleimide, a maleimide derivative of pyrene covalently linked to the SH-groups of Ca-ATPase can also form excimers, which testifies to the formation of Ca-ATPase oligomeric complexes in the native membrane. The probability of oligomerization increases with a thermoinduced decrease in membrane microviscosity, which is correlated with the lowered efficiency of SR Ca-pump operation.
D000818:Animals; D001692:Biological Transport; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002463:Cell Membrane Permeability; D005456:Fluorescent Dyes; D006358:Hot Temperature; D066298:In Vitro Techniques; D009132:Muscles / Q000201:enzymology / Q000378:metabolism*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism*
D005456:Fluorescent Dyes; D000001:Calcimycin; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D002118:Calcium
Mekhanizm termoindutsirovannogo funktsional'nogo razobshcheniia Ca-nasosa sarkoplazmaticheskogo retikuluma skeletnykh myshts.