Involvement of thiol transferase- and thioredoxin-dependent systems in the protection of 'essential' thiol groups of ornithine decarboxylase.
Ornithine decarboxylase (ODC), an enzyme with 'essential' thiol group(s), may be inactivated in vitro by removal of thiol reducing agents and re-activated by soluble factors from rat liver in the presence of NADPH or GSH. The NADPH- and GSH-dependent reducing systems were separated and resolved into three components, called factors A, B1 and B2, by chromatographic techniques. Factor B1 (Mr 12,000) could reactivate ODC in the presence of GSH and co-purified with thiol transferase activity. Factor B2 (Mr 12,000) and factor A (Mr approx. 110,000) were both needed to re-activate ODC in the presence of NADPH, and co-purified with thioredoxin and thioredoxin reductase activity respectively. In an attempt to investigate the physiological role of the 'essential' thiol group(s) of ODC, erythroleukaemia cells were incubated with NN-bis-(2-chloroethyl)-N'-nitrosourea, t-butyl hydroperoxide and vinblastine, which are known to increase the cellular GSSG/GSH ratio, azelaic acid, an inhibitor of thioredoxin reductase, and sodium arsenite, a strong inhibitor of the ODC-re-activating factors. All these compounds were able to decrease significantly the ODC activity induced in these cells. These results suggest that the thiol transferase- and thioredoxin-dependent systems may be physiologically relevant in maintaining ODC in the active, reduced, state.
Flamigni
F
F
Dipartimento di Biochimica, Università di Bologna, Italy.
Marmiroli
S
S
Caldarera
C M
CM
Guarnieri
C
C
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D004793:Enzyme Reactivators / Q000037:antagonists & inhibitors / Q000378:metabolism; D054477:Glutaredoxins; D009955:Ornithine Decarboxylase / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D011490:Protein Disulfide Reductase (Glutathione); D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000201:enzymology
D001426:Bacterial Proteins; D004793:Enzyme Reactivators; D054477:Glutaredoxins; D013438:Sulfhydryl Compounds; D013879:Thioredoxins; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione); D009955:Ornithine Decarboxylase