Abstract
Human erythrocyte (RBC) catechol-O-methyltransferase (COMT) is under genetic control. Experiments were performed to determine whether COMT in the human lymphocyte is regulated in parallel with RBC COMT. Supernatants of lymphocyte homogenates contained COMT activity. However, they also contained a potent COMT inhibitor, the effect of which could be negated by dilution. Lymphocyte COMT activity was maximal at a reaction pH of 7.7 and at a MgCl2 concentration of 0.67mm. The apparent K m value for 3,4-dihydroxybenzoic acid, the catechol substrate for the reaction, was 1.2×10−5 m and that for S-adenosyl-l-methionine, the methyl donor, was 2.3×10−6 m. An average of 48.3±3.3% (mean ± SEM) of the enzyme activity in crude lymphocyte homogenates from 3 subjects was removed by centrifugation at 100,000 g for 1 hr and was presumed to be membrane associated. The average COMT activity in lymphocytes isolated from blood of 23 randomly selected adult subjects was 14.0±1.2 units/106 cells (mean ± SEM) or 913±69 units/mg protein. There was a significant correlation of relative RBC with relative lymphocyte COMT activity in these 23 subjects. The correlation coefficient was 0.733 (P<0.001) when lymphocyte enzyme activity was expressed per milligram of protein and 0.649 (P<0.001) when lymphocyte activity was expressed per 106 cells. These results are compatible with the conclusion that the genetic polymorphism which regulates RBC COMT activity may also regulate the level of human lymphocyte COMT activity.
Similar content being viewed by others
References
Aprille, J. R., and Malamud, D. F. (1975). Catechol-O-methyltransferase in mouse liver plasma membranes. Biochem. Biophys. Res. Commun. 641293.
Axelrod, J., and Cohn, C. K. (1971). Methyltransferase enzymes in red blood cells. J. Pharmacol. Exp. Ther. 176650.
Axelrod, J., and Tomchick, R. (1958). Enzymatic O-methylation of epinephrine and other catechols. J. Biol. Chem. 233702.
Boyum, A. (1968). Separation of leucocytes from blood and bone marrow. Scand. J. Clin. Lab. Invest. (Suppl. 97 211.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72248.
Bremer, J., and Greenberg, D. M. (1961). Enzymatic methylation of foreign sulfhydryl compounds. Biochim. Biophys. Acta 46217.
Cleland, W. W. (1963). Computer programmes for processing enzyme kinetic data. Nature 198463.
De Luca, G., Barberi, I., Ruggeri, P., and DiGiorgio, R. M. (1976). Catechol-O-methyl transferase activity in the individual human platelet populations. Ital. J. Biochem. 25213.
Diliberto, E. J., and Axelrod, J. (1974). Characterization and substrate specificity of a protein carboxymethylase in the pituitary gland. Proc. Natl. Acad. Sci. USA 711701.
Eisenthal, R., and Cornish-Bowden, A. (1974). The direct linear plot. Biochem. J. 139715.
Favour, C. B. (1964). Antigen-antibody reactions in tissue culture. In Ackroyd, J. F. (ed.), Immunological Methods Blackwell Scientific, Oxford, pp. 195–223.
Gershon, E. S., and Jonas, W. Z. (1975). Erythrocyte soluble catechol-O-methyltransferase activity in primary affective disorder. Arch. Gen. Psychiat. 321351.
Giller, E. L., Jr., Young, J. G., Breakefield, X. O., Carbonari, C., Braverman, M., and Cohen, D. J. (1980). Monoamine oxidase and catechol-O-methyltransferase activities in cultured fibroblasts and blood cells from children with autism and Gilles de la Tourette syndrome. Psychiatr. Res. 2187.
Goldstein, D. J., Weinshilboum, R. M., Dunnette, J. H., and Creveling, C. R. (1980). Developmental patterns of catechol-O-methyltransferase in genetically different rat strains: Enzymatic and immunochemical studies. J. Neurochem. 34153.
Groshong, R., Gibson, D. A., and Baldessarini, R. J. (1977). Monoamine oxidase activity in cultured human skin fibroblasts. Clin. Chim. Acta 80113.
Macaione, S., De Luca, G., Barberi, I., and DiGiorgio, R. M. (1976). Methionine activating enzyme in various human platelet populations. Ital. J. Biochem. 25320.
Perper, R. J., Zee, T. W., and Mickelson, M. M. (1968). Purification of lymphocytes and platelets by gradient centrifugation. J. Lab. Clin. Med. 72842.
Poitou, R., and Bohuon, C. (1974). Soluble and membrane catechol-O-methyltransferases in red blood cells of schizophrenic patients. Biomedicine 2191.
Quiram, D. R., and Weinshilboum, R. M. (1976). Inhibition of rat liver catechol-O-methyltransferase by lanthanum, neodymium and europium. Biochem. Pharmacol. 251727.
Raymond, F. A., and Weinshilboum, R. M. (1975). Microassay of human erythrocyte catechol-O-methyltransferase: Removal of inhibitory calcium ion with chelating resin. Clin. Chim. Acta 58185.
Reilly, D. K., Rivera-Calimlim, L., and Van Dyke, D. (1980). Catechol-O-methyltransferase activity: A determinant of levodopa response. Clin. Pharmacol. Ther. 28278.
Roth, J. A. (1980). Presence of membrane-bound catechol-O-methyltransferase in human brain. Biochem. Pharmacol. 293119.
Scanlon, P. D., Raymond, F. A., and Weinshilboum, R. M. (1979). Catechol-O-methyltransferase: Thermolabile enzyme in erythrocytes of subjects homozygous for the allele for low activity. Science 20363.
Spielman, R. S., and Weinshilboum, R. M. (1979). Family studies of low red cell COMT activity. Am. J. Hum. Genet. 31:63A.
Stramentinoli, G., Gualano, M., Algeri, S., de Gaetano, G., and Rossi, E. C. (1978). Catechol-O-methyl transferase (COMT) in human and rat platelets. Thrombosis Haemostasis 39238.
Weinshilboum, R. M. (1978). Human erythrocyte catechol-O-methyltransferase: Correlation with lung and kidney activity. Life Sci. 22625.
Weinshilboum, R. M. (1979). Genetic regulation of catechol-O-methyltransferase. In Gerrendelli, J. A., and Gurvitch, G. (eds), Society for Neuroscience Symposia, Vol. IV, Aspects of Developmental Neurobiology Society for Neuroscience, Washington, D.C., pp. 67–82.
Weinshilboum, R. M., and Raymond, F. A. (1976). Calcium inhibition of rat liver catechol-O-methyltransferase. Biochem. Pharmacol. 25573.
Weinshilboum, R. M., and Raymond, F. A. (1977a). Inheritance of low erythrocyte catechol-O-methyltransferase activity in man. Am. J. Hum. Genet. 29125.
Weinshilboum, R., and Raymond, F. (1977b). Variations in catechol-O-methyltransferase activity in inbred strains of rats. Neuropharmacology 16703.
Weinshilboum, R. M., Raymond, F. A., Elveback, L. R., and Weidman, W. H. (1974). Correlation of erythrocyte catechol-O-methyltransferase activity between siblings. Nature 252490.
Weinshilboum, R. M., Sladek, S., and Klumpp, S. (1979a). Human erythrocyte thiol methyltransferase: Radiochemical microassay and biochemical properties. Clin. Chim. Acta 9759.
Weinshilboum, R. M., Raymond, F. A., and Frohnauer, M. (1979b). Monogenic inheritance of catechol-O-methyltransferase in the rat: Biochemical and genetic studies. Biochem. Pharmacol. 281239.
Wilkinson, G. N. (1961). Statistical estimations in enzyme kinetics. Biochem. J. 80324.
Author information
Authors and Affiliations
Additional information
This study was supported in part by NIH Grants NS 11014, HL 17487, and GM 28157. Dr. Weinshilboum is an Established Investigator of the American Heart Association and a Burroughs Wellcome Scholar in Clinical Pharmacology.
Rights and permissions
About this article
Cite this article
Sladek-Chelgren, S., Weinshilboum, R.M. Catechol-O-methyltransferase biochemical genetics: Human lymphocyte enzyme. Biochem Genet 19, 1037–1053 (1981). https://doi.org/10.1007/BF00484563
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00484563