The kinetic properties and reaction mechanism of histamine methyltransferase from human skin.
The substrate kinetic properties of histamine methyltransferase from human skin were studied at limiting concentrations of both histamine and S-adenosylmethionine. Substrate inhibition by histamine was observed at concentrations above 10 microM. Primary plots showed evidence of a sequential reaction mechanism. The Michaelis constants were derived from secondary plots of slopes from the primary plots ([S]/v versus [S]) versus reciprocal of the second substrate concentration. The mean Km values for histamine and S-adenosylmethionine were 4.2 and 1.8 microM respectively. Histamine in concentrations of 25-100 microM inhibited enzyme activity uncompetitively with respect to S-adenosylmethionine. No substrate inhibition was observed with S-adenosylmethionine. To elucidate the reaction mechanism further, inhibition by the two products, S-adenosylhomocysteine and 1-methylhistamine, was studied. S-Adenosylhomocysteine inhibited non-competitively with respect to histamine and competitively with respect to S-adenosylmethionine. 1-Methylhistamine inhibited non-competitively with respect to histamine and to S-adenosylmethionine. These results are interpreted as providing evidence for an ordered sequential Bi Bi reaction mechanism, with the methyl-group donor S-adenosylmethionine as the first substrate that adds to the enzyme and histamine as the second substrate. 1-Methylhistamine is the first product to leave the enzyme and S-adenosylhomocysteine is the second. The results are discussed in terms of the possible role that this enzyme could play in the modulation of histamine-mediated reactions in skin.
D006632:Histamine / Q000378:metabolism; D006637:Histamine N-Methyltransferase / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008780:Methyltransferases / Q000378:metabolism*; D011485:Protein Binding; D012436:S-Adenosylmethionine / Q000378:metabolism; D012867:Skin / Q000201:enzymology*
D012436:S-Adenosylmethionine; D006632:Histamine; D008780:Methyltransferases; D006637:Histamine N-Methyltransferase