Purification and characterization of antizyme inhibitor of ornithine decarboxylase from rat liver.
A protein inhibiting a protein inhibitor (antizyme) to ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) (ODC), antizyme inhibitor, was purified from the liver cytosol of thioacetamide-treated rats by procedures including antizyme affinity chromatography. Overall purification was roughly estimated to be about 17,000,000-fold and recovery was about 2.4%. The purified preparation showed one major protein band and a faint band corresponding in mobility to molecular weights of 51,000 and 53,500, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Judging from the ornithine decarboxylase activity of the final preparation, the faint band may be ornithine decarboxylase. The apparent molecular weight of antizyme inhibitor estimated by gel filtration on Sephacryl S-200 was approx. 62,000, indicating that antizyme inhibitor may be composed of a single polypeptide chain. In order to examine the question of whether antizyme inhibitor is a protein derived from ornithine decarboxylase, an inactive ornithine decarboxylase, in an immunotitration study and analysis of the binding to antizyme were investigated. The results indicate that antizyme inhibitor may be a protein distinct from ornithine decarboxylase.
Kitani
T
T
Department of Biochemistry, Asahikawa Medical College, Japan.
Fujisawa
H
H
D000818:Animals; D000906:Antibodies / Q000032:analysis; D001665:Binding Sites; D002352:Carrier Proteins; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D065108:Ornithine Decarboxylase Inhibitors; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification*; D011733:Pyridoxamine; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D002352:Carrier Proteins; D065108:Ornithine Decarboxylase Inhibitors; D011506:Proteins; C031017:ornithine decarboxylase antizyme; C039317:ornithine decarboxylase antizyme inhibitor; D011733:Pyridoxamine