EurekaMag.com logo
+ Site Statistics
References:
47,893,527
Abstracts:
28,296,643
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells


, : Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells. Proceedings of the National Academy of Sciences of the United States of America 88(23): 10480-4

The particulate enzyme responsible for the synthesis of endothelium-derived relaxing factor has been purified from cultured and native (noncultured) bovine aortic endothelial cells. Purification of the solubilized particulate enzyme preparation by affinity chromatography on adenosine 2',5'-bisphosphate coupled to Sepharose followed by Superose 6 gel filtration chromatography resulted in a single protein band after denaturing polyacrylamide gel electrophoresis that corresponded to approximately 135 kDa. The enzyme activity in the various fractions was assayed by its stimulatory effect on soluble guanylyl cyclase of rat fetal lung fibroblasts (RFL-6 cells), by the formation of L-citrulline from L-arginine, by measuring nitrite/nitrate formation, and by bioassay on endothelium-denuded vascular strips. Endothelium-derived relaxing factor synthase was purified 3419-fold from the crude particulate fraction of cultured bovine aortic endothelial cells with a 12% recovery (RFL-6 assay). Purified endothelium-derived relaxing factor synthase required L-arginine, NADPH, Ca2+, calmodulin, and 5,6,7,8-tetrahydrobiopterin for full activity.


Accession: 041135330

PMID: 1720542

Submit PDF Full Text: Here


Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:
Close
Close

Related references

Förstermann, U.; Pollock, J.S.; Schmidt, H.H.; Heller, M.; Murad, F., 1991: Calmodulin-dependent endothelium-derived relaxing factor/nitric oxide synthase activity is present in the particulate and cytosolic fractions of bovine aortic endothelial cells. Endothelium-derived relaxing factor/nitric oxide (EDRF/NO) synthesized by bovine aortic endothelial cells and subcellular fractions thereof was assayed by its stimulating effect on soluble guanylyl cyclase of rat fetal lung fibroblasts (RFL-6 cell...

Murray, J.J.; Fridovich, I.; Makhoul, R.G.; Hagen, P.O., 1986: Stabilization and partial characterization of endothelium-derived relaxing factor from cultured bovine aortic endothelial cells. The extreme lability of the chemically undefined vasoregulatory mediator, endothelium-derived relaxing factor (EDRF), has been overcome. The activity of EDRF, from cultured bovine aortic endothelial cells stimulated with A23187, was stabilized by...

Mitchell J.A.; Hecker M.; Sessa W.C.; Swierkosz T.A.; Vane J.R., 1990: L glutamine inhibits the generation of l arginine and the release of endothelium derived relaxing factor edrf from bovine aortic cultured endothelial cells. Journal of Physiology (Cambridge) 429: 73P

Smith, J.A.; Lang, D., 1990: Release of endothelium-derived relaxing factor from pig cultured aortic endothelial cells, as assessed by changes in endothelial cell cyclic GMP content, is inhibited by a phorbol ester. 1. Cultured aortic endothelial cells of the pig respond to the endothelium-derived relaxing factor (EDRF) they release with an increase in cyclic GMP content. This response is inhibited by haemoglobin or by L-NG-monomethyl-arginine (L-NMMA), and h...

Angus, J.A.; Cocks, T.M., 1987: The half-life of endothelium-derived relaxing factor released from bovine aortic endothelial cells in culture. 1. The half-life of endothelium-derived relaxing factor (EDRF) in Krebs solution was determined by bioassay in vitro. 2. A column of bovine aortic endothelial cells grown on microcarrier beads in suspension culture was perfused with Krebs solution...

Wright T.F.; Myers P.R.; Adams H.R., 1991: The effects of endotoxin on endothelium derived relaxing factor and nitric oxide production in cultured aortic endothelial cells. FASEB Journal 5(6): A1728

de Nucci, G.; Gryglewski, R.J.; Warner, T.D.; Vane, J.R., 1988: Receptor-mediated release of endothelium-derived relaxing factor and prostacyclin from bovine aortic endothelial cells is coupled. Bovine aortic endothelial cells were grown on microcarrier beads and were perfused with Krebs-Ringer solution. Endothelium-derived relaxing factor (EDRF) was bioassayed on a cascade of four strips of rabbit aorta, and prostacyclin was analyzed by...

D'Orléans-Juste, P.; de Nucci, G.; Vane, J.R., 1989: Kinins act on B1 or B2 receptors to release conjointly endothelium-derived relaxing factor and prostacyclin from bovine aortic endothelial cells. 1. Bradykinin (Bk) induced the coupled release of endothelium-derived relaxing factor (EDRF) and prostacyclin (PGI2) from bovine aortic endothelial cells grown in culture. The B2 kinin receptor antagonist, [D-Arg0,Hyp3,Thi5,8,D-Phe7]-Bk, abolished...

Forstermann U.; Pollock J.S.; Schmidt H.H.H.W.; Mitchell J.A.; Kohlhaas K.L.; Heller M.; Murad F., 1991: Characterization and purification of particulate edrf synthase from bovine aortic endothelial cells. FASEB Journal 5(6): A1728

Kuhn, M.; Otten, A.; Frölich, J.C.; Förstermann, U., 1991: Endothelial cyclic GMP and cyclic AMP do not regulate the release of endothelium-derived relaxing factor/nitric oxide from bovine aortic endothelial cells. The present study was designed to investigate whether cyclic GMP or cyclic AMP modulates the release of endothelium-derived relaxing factor/nitric oxide (EDRF/NO) in cultured bovine aortic endothelial cells (BAE cells). BAE cell-conditioned medium...