Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. 1992

K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
Cardiology Division, Emory University School of Medicine, Atlanta, Georgia 30322.

The constitutive endothelial cell nitric oxide synthase (NOS) importantly regulates vascular homeostasis. To gain understanding of this enzyme, a pEF BOS cDNA library of 5 x 10(5) clones was prepared from bovine aortic endothelial cells (BAEC) and screened with a 2.8-kb cDNA BamHI fragment of rat brain NOS. Clone pBOS13 was found to express NO synthase activity when transfected into COS-7 cells. Sequence analysis revealed sequences compatible with binding domains for calcium/calmodulin, flavin mononucleotide, flavin adenine nucleotide and NADPH. The deduced amino acid sequence revealed a protein with a relative mol mass of 133,286, which is 58% homologous to the rat cerebellar NOS and 51% homologous to the mouse macrophage NOS. The amino-terminal portion of the protein exhibits several characteristics peculiar to the endothelial cell NOS. These include a proline-rich region and several potential sites for proline-directed phosphorylation as well as a potential substrate site for acyl transferase. Northern hybridization to mRNA from cultured BAEC revealed an abundant 4.8-kb message, which was not increased by coincubation with tumor necrosis factor alpha, but was markedly increased by exposure to shear stress for 24 h. The unique features of the endothelial cell NO synthase, particularly in the amino terminal portion of the molecule, may provide for novel regulatory influences of enzyme activity and localization.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D002417 Cattle Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor. Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004730 Endothelium, Vascular Single pavement layer of cells which line the luminal surface of the entire vascular system and regulate the transport of macromolecules and blood components. Capillary Endothelium,Vascular Endothelium,Capillary Endotheliums,Endothelium, Capillary,Endotheliums, Capillary,Endotheliums, Vascular,Vascular Endotheliums
D000594 Amino Acid Oxidoreductases A class of enzymes that catalyze oxidation-reduction reactions of amino acids. Acid Oxidoreductases, Amino,Oxidoreductases, Amino Acid
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001011 Aorta The main trunk of the systemic arteries. Aortas
D015971 Gene Expression Regulation, Enzymologic Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis. Enzymologic Gene Expression Regulation,Regulation of Gene Expression, Enzymologic,Regulation, Gene Expression, Enzymologic

Related Publications

K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
August 1996, Biochimica et biophysica acta,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
July 1992, Proceedings of the National Academy of Sciences of the United States of America,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
August 1992, FEBS letters,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
May 1996, European journal of biochemistry,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
December 1995, The Journal of clinical investigation,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
January 1992, Journal of cardiovascular pharmacology,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
April 2000, Acta pharmacologica Sinica,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
August 1992, The Journal of biological chemistry,
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
September 1999, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.),
K Nishida, and D G Harrison, and J P Navas, and A A Fisher, and S P Dockery, and M Uematsu, and R M Nerem, and R W Alexander, and T J Murphy
January 2002, Methods in enzymology,
Copied contents to your clipboard!