Isolation of the low-molecular-mass form of catechol O-methyltransferase from rat liver and immunocytochemical localization of the enzyme in the glycogen compartment.
The low-molecular-mass form of two distinct catechol O-methyltransferase activities (S-adenosyl-L-methionine: catechol O-methyltransferase, COMT, EC 2.1.1.6) has been purified to homogeneity from rat liver using 40-70% ammonium sulfate precipitation, gel filtration on Sephadex G-100, adsorption on hydroxyapatite C and ion-exchange chromatography on DEAE-Sepharose CL-6B. The relative molecular mass Mr, determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis is 22 400 +/- 500. Irradiation of the enzyme in the presence of 8-azido-[methyl-3H]AdoMet results in the specific labeling of the catalytic site of the enzyme. Photolabeling was successful with crude COMT preparations and with the isolated enzyme. Immunocytochemical studies present new information about the localization of the low-molecular-mass form in the liver parenchyma. Subcellularly COMT immunoreactivity could be attributed exclusively to the compartment with glycogen granules. Nucleus, mitochondria and endoplasmic reticulum showed no immunostaining.
D000918:Antibody Specificity; D002394:Catechol O-Methyltransferase / Q000276:immunology / Q000302:isolation & purification*; D002451:Cell Compartmentation; D007124:Immunoenzyme Techniques; D008099:Liver / Q000378:metabolism*; D008112:Liver Glycogen / Q000378:metabolism*; D008854:Microscopy, Electron
D008112:Liver Glycogen; D002394:Catechol O-Methyltransferase