Inducible nitric oxide synthase from human articular chondrocytes: cDNA cloning and analysis of mRNA expression

Biochim Biophys Acta. 1994 Sep 21;1208(1):145-50. doi: 10.1016/0167-4838(94)90171-6.


Human articular chondrocytes can be induced by IL-1 beta, TNF-alpha or LPS to release high levels of nitric oxide. Using degenerate PCR primers based on homologous regions from previously cloned NOS enzymes, a 1.9 kb cDNA fragment was amplified from IL-1 beta stimulated but not from resting chondrocytes. Screening of a lambda gt11 cDNA library, which was prepared from RNA of IL-1 beta activated chondrocytes, resulted in the isolation of the complete cDNA, encoding a protein of 1153 amino acids. Comparison of the cDNA sequence identified human chondrocyte iNOS to be almost identical to the sequence recently reported for the hepatocyte enzyme, differing in 12 amino acids. Northern blot analysis revealed, that stimulated chondrocytes express a single 4.5 kb iNOS mRNA species. IL-1 beta induction of iNOS mRNA was detectable by 6 h and continued to be elevated throughout a 72 h culture period. Screening of a human bone cDNA library identified this inducible NOS to be also expressed by bone cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Oxidoreductases / biosynthesis*
  • Amino Acid Oxidoreductases / chemistry
  • Amino Acid Oxidoreductases / genetics
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Blotting, Northern
  • Cartilage, Articular / enzymology*
  • Cloning, Molecular*
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics*
  • Gene Expression*
  • Humans
  • In Situ Hybridization
  • Interleukin-1 / pharmacology
  • Molecular Sequence Data
  • Nitric Oxide Synthase
  • Polymerase Chain Reaction
  • RNA, Messenger / analysis*
  • RNA, Messenger / metabolism


  • DNA, Complementary
  • Interleukin-1
  • RNA, Messenger
  • Nitric Oxide Synthase
  • Amino Acid Oxidoreductases

Associated data

  • GENBANK/U05810