Purification and cDNA sequence of an inducible nitric oxide synthase from a human tumor cell line.
A combination of cytokines induced the expression of nitric oxide synthase (NOS) in a human colorectal adenocarcinoma cell line, DLD-1. We have purified the enzyme and examined some of its biochemical properties. An antiserum to an inducible NOS from murine macrophages cross-reacted with the DLD-1 NOS. The purified human and murine enzymes displayed a similar lack of dependence on exogenous calcium and calmodulin for activity, which contrasts with the requirement for calcium and calmodulin of purified brain and endothelial isoforms of NOS. We have also isolated a cDNA for a cytokine-induced NOS from DLD-1 cells. Sequence analysis of this cDNA and NOS cDNAs from human liver, smooth muscle, and macrophages suggests that, at the genetic level, there is a single isoform of human-inducible NOS.
Sherman
P A
PA
Division of Experimental Therapy, Burroughs Wellcome Company, Research Triangle Park, North Carolina 27709.
Laubach
V E
VE
Reep
B R
BR
Wood
E R
ER
D000230:Adenocarcinoma / Q000201:enzymology; D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015179:Colorectal Neoplasms / Q000201:enzymology; D016207:Cytokines / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry; D004790:Enzyme Induction; D006801:Humans; D008264:Macrophages / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D016207:Cytokines; D018076:DNA, Complementary; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases