Properties of a partially purified phosphodimethylethanolamine methyltransferase from rat brain cytosol.
The presence of cytosolic S-adenosylmethionine-dependent N-methyltransferase(s) activity(ies) capable of converting phosphoethanolamine into phosphocholine has been recently demonstrated in the rat brain. At least two enzymes are involved in the methylation of phosphoethanolamine to phosphocholine and these are separable by ammonium sulphate fractionation. The enzyme catalysing the last step of this methylation process is present in the 50-80% ammonium sulphate fraction. A 220-fold purified enzyme has been obtained with sequentially employed Q-Sepharose fast flow and octyl-Sepharose CL4B column chromatography. The maximum enzyme activity was at pH 9.5. The Km values for S-adenosylmethionine, the methyl donor, and phosphodimethylethanolamine, the methyl acceptor, were 125 microM and 750 microM respectively. This phosphodimethylethanolamine N-methyltransferase was found to be calcium-dependent, with a 4-fold increase in activity at 0.5 mM-CaCl2. S-Adenosylhomocysteine at 0.5 mM caused 100% inhibition of the activity. The effects of various structural analogues on the phosphodimethylethanolamine N-methyltransferase activity were also investigated and these results suggest that the enzyme is specific to the substrate. These results provide evidence for the existence of the pathway for the methylation of phosphoethanolamine to phosphocholine in rat brain cytosol.
Andriamampandry
C
C
Department of Biochemistry and Molecular Biology, University of Manitoba, Winnipeg, Canada.
Massarelli
R
R
Kanfer
J N
JN
D000645:Ammonium Sulfate; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004983:Ethanolamines / Q000378:metabolism; D005590:Fractional Precipitation; D006863:Hydrogen-Ion Concentration; D008297:Male; D008745:Methylation; D008780:Methyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D010767:Phosphorylcholine / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012435:S-Adenosylhomocysteine / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000378:metabolism
D004983:Ethanolamines; D010767:Phosphorylcholine; C005448:phosphorylethanolamine; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; C078173:phosphodimethylethanolamine N-methyltransferase; D008780:Methyltransferases; D000645:Ammonium Sulfate; D002118:Calcium