title: Partial purification of a phosphoethanolamine methyltransferase from rat brain cytosol.
abstract: The conversion of phosphoethanolamine to phosphocholine requires 3 separate N-methyltransferases. We had previously purified the enzyme catalyzing the last methylation, phosphodimethylethanolamine N-methyltransferase. We have successfully purified the enzyme catalyzing the initial methylation of phosphoethanolamine. A 434 fold purified enzyme from rat brain was obtained by the sequential use of ammonium sulfate fractionation, Q-Sepharose fast flow column chromatography and a omega-aminoethyl agarose column chromatography. The pH optimum was 11 or greater, the Km value for phosphoethanolamine was 167.8 +/- 41.7 microM and the Vmax was 487.3 +/- 85 mmoles/mg/hr. The kinetics for S-adenosyl-methionine, the methyldonor, has characteristics of cooperative binding with a Km of 1.805 +/- 0.59 mM and a Vmax of 16.9 +/- 3.6 mumoles/mg/hr. The activity was stimulated 6 fold by 2.5 mM MnCl2 and inhibited by DZA and S-adenosylhomocysteine. These results reinforce the early in vivo observations which had provided suggestive evidence for the existence of a pathway for the methylation of phosphoethanolamine to phosphocholine in rat brain.
authors: University of Manitoba, Winnipeg, Canada.
authors: S
authors: S
authors: Mukherjee
authors: L
authors: L
authors: Freysz
authors: J N
authors: JN
authors: Kanfer
mesh_terms: D000645:Ammonium Sulfate; D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D002413:Cations, Divalent; D011232:Chemical Precipitation; D002847:Chromatography, Agarose; D003600:Cytosol / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008780:Methyltransferases / Q000302:isolation & purification*; D051381:Rats; D012436:S-Adenosylmethionine
chemical_list: D002413:Cations, Divalent; D012436:S-Adenosylmethionine; D008780:Methyltransferases; C102844:phosphoethanolamine methyltransferase; D000645:Ammonium Sulfate