Purification and immunochemical characterization of monoamine oxidase from rat and human liver.
1. Monoamine oxidase from rat and human liver was purified to homogeneity by the criterion of polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. 2. The enzyme activity was extracted from mitochondrial preparations by Triton X-100. The enzyme was purified by (NH4)2SO4 fractionation followed by chromatography on DEAE-cellulose, Sepharose 6B, spheroidal hydroxyapatite, and finally chromatography on diazo-coupled tyramine-Sepharose. 3. Distinct differences occur in the chromatographic behaviour of the two enzymes on both DEAE-cellulose and spheroidal hydroxyapatite. 4. It is unlikely that the purification of the enzymes on tyramine-Sepharose is due to affinity chromatography and reasons for this are discussed. 5. The purified enzymes did not oxidize-5-hydroxytryptamine and the relative activities of the enzymes with benzylamine were increased approx. 1.25-fold compared with the enzyme activities of mitochondrial preparations. 6. Immunotitration of enzyme activity in extracts of mitochondrial preparations from rat liver was carried out with 5-hydroxytryptamine, tyramine and benzylamine. The enzyme activities were completely immunoprecipitated by the same volume of antiserum. Similar results were obtained with the antiserum to the enzyme from human liver.
D000818:Animals; D001596:Benzylamines; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D006801:Humans; D006882:Hydroxyapatites; D007118:Immunoassay; D008930:Mitochondria, Liver / Q000201:enzymology*; D008995:Monoamine Oxidase / Q000302:isolation & purification / Q000378:metabolism; D011092:Polyethylene Glycols; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives / Q000138:chemical synthesis; D012701:Serotonin; D014439:Tyramine
D001596:Benzylamines; D006882:Hydroxyapatites; D012701:Serotonin; D011092:Polyethylene Glycols; D012685:Sepharose; D008995:Monoamine Oxidase; D014439:Tyramine