Production of an EDRF-like activity in the cytosol of N1E-115 neuroblastoma cells.
Accumulation of cyclic GMP in cultured rat lung fibroblasts was used to test the hypothesis that N1E-115 neuroblastoma cells produce an endothelium-derived relaxing factor (EDRF)-like activity. By using this assay, the production of an EDRF-like activity in homogenates and cytosolic fractions of N1E-115 neuroblastoma cells was observed. Detection of the activity required the presence of superoxide dismutase and was inhibited by hemoglobin. Production of the EDRF-like factor was dependent on L-arginine and NADPH. The apparent Km for L-arginine was 1.25 microM and the apparent Km for NADPH was 1.67 microM. The production of the EDRF-like activity was inhibited by the L-arginine analogs, NG-monomethyl-L-arginine and NG-nitro-L-arginine, with apparent Ki values of 1.0 and 0.3 microM, respectively.
Gorsky
L D
LD
Pharmaceutical Discovery, Abbott Laboratories, Illinois 60064.
Förstermann
U
U
Ishii
K
K
Murad
F
F
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002474:Cell-Free System / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000378:metabolism*; D019335:Nitroarginine; D014407:Tumor Cells, Cultured / Q000378:metabolism*; D019323:omega-N-Methylarginine
D019335:Nitroarginine; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D001120:Arginine; D006152:Cyclic GMP