Redox-dependent and redox-independent subcomponents of protein degradation in perfused myocardium.
The integration of proteolytic pathways with metabolism was investigated in perfused rat myocardium. After a 10-min incorporation period, the minute-to-minute release of [3H]leucine from myocardial proteins was measured in nonrecirculating effluent perfusate. The nontoxic pro-oxidant probe diamide (100 microM) or a supraphysiological concentration of the endogenous oxidative metabolite dehydroascorbic acid (200 microM) reversibly inhibited 75% of myocardial proteolysis consisting of several known subcomponents (redox dependent); however, 25% of proteolysis was diamide insensitive (redox independent). Decrease in extracellular glucose concentration from 10 to 0.1 mM strongly increased the potencies of minimally effective concentrations of diamide (10 microM) or dehydroascorbic acid (15 microM) by approximately 10-fold to the respective potencies maximally inhibiting proteolysis. The reversal of diamide action was also strongly dependent on the perfusate glucose concentration observed at 0.1, 0.2, 1.0 and 10 mM glucose. Proteolytic inhibition caused by diamide (100 microM) was not accompanied by change in basal tissue ATP content of 5 micromol/g wet wt. Conversely, nearly lethal 60% ATP depletion caused by sodium azide (0.4 mM) was not accompanied by change in total [3H]leucine release. Results indicate that a large proteolytic subcomponent (75%) is maintained by redox chains fed by glucose; however, there is no apparent linkage of this proteolysis to short-term ATP fluctuations. A distinct major proteolytic subcomponent (25%) does not vary in response to experimental intervention in either ATP content or redox chains.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003683:Dehydroascorbic Acid / Q000008:administration & dosage / Q000494:pharmacology; D003958:Diamide / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D005947:Glucose / Q000008:administration & dosage / Q000494:pharmacology; D007930:Leucine / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D010477:Perfusion; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D019810:Sodium Azide / Q000494:pharmacology; D014316:Tritium