Endocrinology and Metabolism

Redox-dependent and redox-independent subcomponents of protein degradation in perfused myocardium

Thomas D. Lockwood


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 μM) or a supraphysiological concentration of the endogenous oxidative metabolite dehydroascorbic acid (200 μM) 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 μM) or dehydroascorbic acid (15 μM) by ∼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 μM) was not accompanied by change in basal tissue ATP content of 5 μmol/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.

  • proteolysis
  • oxidation-reduction
  • adenosine 5′-triphosphate
  • diamide
  • sodium azide
  • glucose


  • Address for reprint requests and other correspondence: T. D. Lockwood, Dept. of Pharmacology and Toxicology, School of Medicine, Wright State University, Dayton, OH 45435 (E-Mail:thomas.lockwood{at}wright.edu).

  • The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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