Stimulation of protein methylase II from Torpedo marmorata by cholinergic effectors.
The enzymatic transfer of methyl groups mediated by protein methylase II onto proteins of the electroplaque tissue of Torpedo marmorata is described. The protein methylase II resides to the extent of 80% in the cytosol and 20% in the acetylcholine receptor-rich membrane. The kinetics of the methyl-group transfer are characteristically different in the cytosol and membrane fractions. The reaction is inhibited by phosphate with IC50 = 450 microM. The cholinergic effectors carbamoylcholine, flaxedil and alpha-bungarotoxin applied to the outside of the acetylcholine receptor-rich membrane vesicles stimulated the protein methylase II which is exclusively located inside the vesicles. The stimulation is biphasic and transient, yielding an increased initial velocity and a peak of activity at 2 min after the addition of the effector. The stimulation by carbamoylcholine is qualitatively similar to that elicited by the antagonist. In addition, the protein methylase II is stimulated transiently by phospholipase A2 with a time-course clearly different from that of the cholinergic effectors. We conclude that the conformational change in the receptor-protein elicited by cholinergic effectors is efficiently transduced to the cytoplasmic methylation sites.
D000818:Animals; D002038:Bungarotoxins / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004557:Electric Organ / Q000201:enzymology*; D005703:Gallamine Triethiodide / Q000494:pharmacology; D007700:Kinetics; D008745:Methylation; D010710:Phosphates / Q000494:pharmacology; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D011496:Protein Methyltransferases / Q000378:metabolism*; D011497:Protein O-Methyltransferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000502:physiology*; D012436:S-Adenosylmethionine / Q000378:metabolism; D014101:Torpedo / Q000378:metabolism*
D002038:Bungarotoxins; D010710:Phosphates; D011950:Receptors, Cholinergic; D012436:S-Adenosylmethionine; D002217:Carbachol; D011496:Protein Methyltransferases; D011497:Protein O-Methyltransferase; D010741:Phospholipases A; D054467:Phospholipases A2; D005703:Gallamine Triethiodide