Different sensitivities of native and oxidized forms of Na+/K(+)-ATPase to intracellular proteinases.
Inactivation of Na+/K(+)-ATPase by partially reduced oxygen metabolites has been implicated in ischemia-reperfusion injury to heart and other organs. Because oxidation of many proteins makes them more susceptible to degradation by intracellular proteinases, we studied the effects of several such proteinases on native and H2O2-oxidized preparations of Na+/K(+)-ATPase from canine kidney (containing alpha 1 isoform of the catalytic subunit) and rat axolemma (containing alpha 2 and alpha 3 isoforms). Lysosomal cathepsin D degraded the native and the oxidized preparations at acid pH, but it was significantly more effective against the oxidized forms. m-Calpain had little or no effect on the native Na+/K(+)-ATPase preparations, but it digested the oxidized alpha-subunits of the axolemma and the kidney enzymes. mu-Calpain's effects were similar to those of m-calpain. Multi-catalytic proteinase which is known to degrade a large number of oxidized proteins, did not affect the native or the oxidized forms of Na+/K(+)-ATPase. The findings suggest that (a) during oxidative stress there may be accelerated degradation of the oxidatively damaged Na+/K(+)-ATPase, either through internalization and transport to lysosomes, or by the action of calpains at the membrane; and (b) those isoforms of the enzyme that are more sensitive to oxidants are more susceptible to degradation by the above processes.
Zolotarjova
N
N
Department of Pharmacology, Medical College of Ohio, Toledo 43699-0008.
Ho
C
C
Mellgren
R L
RL
Askari
A
A
Huang
W H
WH
D000818:Animals; D002154:Calpain / Q000494:pharmacology; D002402:Cathepsin D / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000494:pharmacology; D004285:Dogs; D010450:Endopeptidases / Q000378:metabolism* / Q000494:pharmacology; D006861:Hydrogen Peroxide; D007679:Kidney Medulla / Q000201:enzymology; D009097:Multienzyme Complexes / Q000494:pharmacology; D010084:Oxidation-Reduction; D046988:Proteasome Endopeptidase Complex; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D009097:Multienzyme Complexes; D006861:Hydrogen Peroxide; D010450:Endopeptidases; D002154:Calpain; D003546:Cysteine Endopeptidases; D002402:Cathepsin D; D046988:Proteasome Endopeptidase Complex; D000254:Sodium-Potassium-Exchanging ATPase