Light activation of phosphatidylethanolamine N-methyltransferase in rod outer segments and its modulation by association states of transducin.
Phosphatidylethanolamine N-Methyltransferase (PE N-MTase) is the enzyme responsible for the synthesis of phosphatidylcholine from phosphatidylethanolamine by successive transfer of methyl groups. This enzyme is present in bovine rod outer segments (ROS) and it is the only pathway for the synthesis of phosphatidylcholine in the outer segment of rod photoreceptor cells. In dark-adapted ROS membranes PE N-MTase activity is stimulated by 100% when ROS membranes are incubated under light condition. To determine whether the retinal G protein, transducin (Gt), intervenes in the regulation of PE N-MTase in these membranes, the effects of guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) and guanosine 5'-O-(2-thiodiphosphate (GDPbetaS) on the enzyme activity were examined. In dark, GTPgammaS which induces dissociation of Gt, stimulates the enzyme activity mimicking the stimulation by light. On the contrary, GDPbetaS stabilizes the inactive state of Gt, inhibiting the stimulation by light of PE N-MTase without affecting basal activities. In addition, adenosine 5'-diphosphate (ADP)-ribosylation by cholera and pertussis toxin was studied. ADP-ribosylation of ROS membrane with pertussis toxin, which stabilizes transducin in its inactive state, prevents the light-induced increase in PE N-MTase activity. On the contrary ADP-ribosylation with cholera toxin stimulates the enzyme activity. Our findings therefore suggest that light-stimulated effect of PE N-MTase activity is transducin-mediated.
Roque
M E
ME
Instituto de Investigaciones Bioquímicas, Universidad Nacional del Sur y Consejo Nacional de Investigaciones Científicas y Técnicas, Bahía Blanca, FWB, B 8000, Argentina.
Salvador
G A
GA
Giusto
N M
NM
D000244:Adenosine Diphosphate / Q000502:physiology; D000818:Animals; D002417:Cattle; D002772:Cholera Toxin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D008029:Lighting; D008780:Methyltransferases / Q000187:drug effects / Q000378:metabolism*; D037342:Pertussis Toxin; D050918:Phosphatidylethanolamine N-Methyltransferase; D010775:Photic Stimulation; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D013873:Thionucleotides / Q000494:pharmacology; D015293:Transducin / Q000502:physiology*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D013873:Thionucleotides; D010566:Virulence Factors, Bordetella; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000244:Adenosine Diphosphate; C023427:guanosine 5'-O-(2-thiodiphosphate); D002772:Cholera Toxin; D008780:Methyltransferases; D050918:Phosphatidylethanolamine N-Methyltransferase; D037342:Pertussis Toxin; D015293:Transducin