Characterisation of nitric oxide synthase activity in the tropical sea anemone Aiptasia pallida.
The presence of nitric oxide synthase (EC 1.14.23 NOS) activity is demonstrated in the tropical marine cnidarian Aiptasia pallida (Verrill). Enzyme activity was assayed by measuring the conversion of [3H]arginine to [3H]citrulline. Optimal NOS activity was found to require NADPH. Activity was inhibited by the competitive NOS inhibitor NG-methyl-L-arginine (L-NMA), but not the arginase inhibitors L-valine and L-ornithine. NOS activity was predominantly cytosolic, and was characterised by a Km for arginine of 19.05 microM and a Vmax of 2.96 pmol/min per microgram protein. Histochemical localisation of NOS activity using NADPH diaphorase staining showed the enzyme to be predominantly present in the epidermal cells and at the extremities of the mesoglea. These results provide a preliminary biochemical characterisation and histochemical localisation of NOS activity in A. pallida, an ecologically important sentinel species in tropical marine ecosystems.
Morrall
C E
CE
Plymouth Environmental Research Centre, University of Plymouth, UK. cmorrall@bbsr.edu
Galloway
T S
TS
Trapido-Rosenthal
H G
HG
Depledge
M H
MH
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002956:Citrulline / Q000032:analysis / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D009252:NADPH Dehydrogenase / Q000032:analysis / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012615:Sea Anemones / Q000201:enzymology* / Q000502:physiology; D013194:Staining and Labeling / Q000379:methods; D014633:Valine / Q000494:pharmacology; D019323:omega-N-Methylarginine
D001120:Arginine; D002956:Citrulline; D004791:Enzyme Inhibitors; D009249:NADP; D009252:NADPH Dehydrogenase; D019331:NG-Nitroarginine Methyl Ester; D019001:Nitric Oxide Synthase; D014633:Valine; C092506:N(G)-monomethylarginine acetate; D019323:omega-N-Methylarginine