Inhibition of glutathione reductase by oncomodulin.
Evidence for a specific interaction between oncomodulin and glutathione reductase is presented. Glutathione reductase (EC 1.6.4.2) isolated from either the bovine intestinal mucosa or the rat liver was bound in a Ca2(+)-dependent manner to oncomodulin which was covalently attached to Sepharose. In addition, glutathione reductase was able to catalyze the reduction of the disulfide-linked dimer of oncomodulin. The interaction of these proteins could also be indirectly demonstrated by monitoring glutathione reductase activity since oncomodulin was shown to inhibit the enzyme in a dose-dependent manner with an apparent IC50 of approximately 5 microM. The kinetic analysis of the oncomodulin-dependent effects on glutathione reductase activity indicates that oncomodulin interacts at a site other than the active site as the oncomodulin-induced inhibition was of the noncompetitive type. The in vivo inhibition of glutathione reductase appears to be an oncomodulin-specific effect as closely related members of the troponin C superfamily such as rabbit (pI 5.5) or carp (pI 4.25) parvalbumins, as well as calmodulin, failed to affect the activity of this enzyme. The present in vitro study indicating that oncomodulin can regulate the activity of glutathione reductase could be very significant with respect to the elucidation of a physiological role for oncomodulin.
Palmer
E J
EJ
Division of Biological Sciences, National Research Council of Canada, Ottawa, Ontario.
MacManus
J P
JP
Mutus
B
B
D000818:Animals; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D005980:Glutathione Reductase / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D007413:Intestinal Mucosa / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008114:Liver Neoplasms, Experimental; D009363:Neoplasm Proteins / Q000494:pharmacology*; D051381:Rats
D002135:Calcium-Binding Proteins; D009363:Neoplasm Proteins; C034733:oncomodulin; D005980:Glutathione Reductase; D005978:Glutathione; D019803:Glutathione Disulfide