Antigen: human eNOS amino acids 1186-1203 (RGAVPWAFDPPGSDTNSP) · Host: rabbit · Application(s): WB, IP; other applications not tested · eNOS catalyzes the formation of NO from L-arginine in many cell types including vascular endothelium, bronchiolar epithelium, cardiac myocytes, spleen, and kidney.^1,2,3 Although eNOS was initially characterized as a constitutive enzyme, expression of eNOS mRNA is altered by many factors including shear stress, fibroblast growth factor, hypoxia, and LPS.^4,5,6 eNOS is myristoylated and palmitoylated at the N-terminus, modifications which are required for localization to the plasmalalemmal caveolae of endothelial cells.^7,8,9,10,11 Additionally, eNOS is phosphorylated on Ser¹¹⁷⁹ by protein kinase Akt resulting in an increase in NOS activity.^12,13,14

1 Janssens, S.P., Shimouchi, A., Quertermous, T., et al. Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase. J Biol Chem 267 14519-14522 (1992).

2 Shaul, P.W., North, A.J., Wu, L.C., et al. Endothelial nitric oxide synthase is expressed in cultured human bronchiolar epithelium. J Clin Invest 94 2231-2236 (1994).

3 Balligand, J., Kobzik, L., Han, X., et al. Nitric oxide-dependent parasympathetic signaling is due to activation of constitutive endothelial (type III) nitric oxide synthase in cardiac myocytes. J Biol Chem 270 14582-14586 (1995).

4 Xiao, Z., Zhang, Z., and Diamond, S.L. Shear stress induction of the endothelial nitric oxide synthase gene is calcium-dependent but not calcium-activated. J Cell Physiol 171 205-211 (1997).

5 Arnett, U.A., McMillan, A., Dinerman, J.L., et al. Regulation of endothelial nitric-oxide synthase during hypoxia. J Biol Chem 271 15069-15073 (1996).

6 Lu, J., Schmiege, L.M., Kuo, L., et al. Downregulation of endothelial constitutive nitric oxide synthase expression by lipopolysaccharide. Biochem Biophys Res Commun 225 1-5 (1996).

7 Busconi, L., and Michel, T. Endothelial nitric oxide synthase. N-terminal myristoylation determines subcellular localization. J Biol Chem 268 8410-8413 (1993).

8 Robinson, L.J., and Michel, T. Mutagenesis of palmitoylation sites in endothelial nitric oxide synthase identifies a novel motif for dual acylation and subcellular targeting. Proc Natl Acad Sci USA 92 11776-11780 (1995).

9 Feron, O., Saldana, F., Michel, J.B., et al. The endothelial nitric-oxide synthase-caveolin regulatory cycle. J Biol Chem 273 3125-3128 (1998).

10 García-Cardeña, G., Oh, P., Liu, J., et al. Targeting of nitric oxide synthase to endothelial cell caveolae via palmitoylation: Implications for nitric oxide signaling. Proc Natl Acad Sci USA 93 6448-6453 (1996).

11 Feron, O., Belhassen, L., Kobzik, L., et al. Endothelial nitric oxide synthase targeting to caveolae. Specific interactions with caveolin isoforms in cardiac myocytes and endothelial cells. J Biol Chem 271 22810-22814 (1996).

12 Corson, M.A., James, N.L., Latta, S.E., et al. Phosphorylation of endothelial nitric oxide synthase in response to fluid shear stress. Circ Res 79 984-991 (1996).

13 Dimmeler, S., Fleming, I., Fisslthaler, B., et al. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 399 601-605 (1999).

14 Fulton, D., Gratton, J., McCabe, T.J., et al. Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 399 597-601 (1999).

Shaul, P.W., North, A.J., Wu, L.C., et al. Endothelial nitric oxide synthase is expressed in cultured human bronchiolar epithelium. J Clin Invest 94 2231-2236 (1994).

Balligand, J., Kobzik, L., Han, X., et al. Nitric oxide-dependent parasympathetic signaling is due to activation of constitutive endothelial (type III) nitric oxide synthase in cardiac myocytes. J Biol Chem 270 14582-14586 (1995).

Nishida, K., Harrison, D.G., Navas, J.P., et al. Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. J Clin Invest 90 2092-2096 (1992).

Janssens, S.P., Shimouchi, A., Quertermous, T., et al. Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase. J Biol Chem 267 14519-14522 (1992).

García-Cardeña, G., Oh, P., Liu, J., et al. Targeting of nitric oxide synthase to endothelial cell caveolae via palmitoylation: Implications for nitric oxide signaling. Proc Natl Acad Sci USA 93 6448-6453 (1996).

Busconi, L., and Michel, T. Endothelial nitric oxide synthase. N-terminal myristoylation determines subcellular localization. J Biol Chem 268 8410-8413 (1993).

Feron, O., Saldana, F., Michel, J.B., et al. The endothelial nitric-oxide synthase-caveolin regulatory cycle. J Biol Chem 273 3125-3128 (1998).

Xiao, Z., Zhang, Z., and Diamond, S.L. Shear stress induction of the endothelial nitric oxide synthase gene is calcium-dependent but not calcium-activated. J Cell Physiol 171 205-211 (1997).

Fulton, D., Gratton, J., McCabe, T.J., et al. Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 399 597-601 (1999).

Feron, O., Belhassen, L., Kobzik, L., et al. Endothelial nitric oxide synthase targeting to caveolae. Specific interactions with caveolin isoforms in cardiac myocytes and endothelial cells. J Biol Chem 271 22810-22814 (1996).

Dimmeler, S., Fleming, I., Fisslthaler, B., et al. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 399 601-605 (1999).

Arnett, U.A., McMillan, A., Dinerman, J.L., et al. Regulation of endothelial nitric-oxide synthase during hypoxia. J Biol Chem 271 15069-15073 (1996).

Robinson, L.J., and Michel, T. Mutagenesis of palmitoylation sites in endothelial nitric oxide synthase identifies a novel motif for dual acylation and subcellular targeting. Proc Natl Acad Sci USA 92 11776-11780 (1995).

Corson, M.A., James, N.L., Latta, S.E., et al. Phosphorylation of endothelial nitric oxide synthase in response to fluid shear stress. Circ Res 79 984-991 (1996).

Lu, J., Schmiege, L.M., Kuo, L., et al. Downregulation of endothelial constitutive nitric oxide synthase expression by lipopolysaccharide. Biochem Biophys Res Commun 225 1-5 (1996).

Gnanapandithen, K., Chen, Z., Kau, C., et al. Cloning and characterization of murine endothelial constitutive nitric oxide synthase. Biochim Biophys Acta 1308 103-106 (1996).

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