Endothelial nitric oxide synthase activity is linked to its presence at cell–cell contacts

Govers, Roland; Bevers, L.E.; Bree, Petra de; Rabelink, Ton J.

doi:10.1042/bj3610193

Cited by 68 publications

(65 citation statements)

References 44 publications

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“…Syndecans that contain both HS and CS have an established association with the cytoskeleton [19], and through it can decentralize the signal by distributing it to multiple sites within the cell (i.e., nucleus, organelles, focal adhesions, intercellular junctions). Significantly, the platelet-endothelial cell adhesion molecule (PECAM-1) associates with the cytoskeleton through catenins, and has been linked to shearinduced eNOS activation [20][21][22]. In terms of central transduction, it is noteworthy that glypicans which contain HS, but not CS, are linked to caveolae where eNOS resides along with many other signaling molecules [3].…”

Section: Discussionmentioning

confidence: 99%

The role of endothelial glycocalyx components in mechanotransduction of fluid shear stress

Pahakis¹,

Kosky²,

Dull³

et al. 2007

Biochemical and Biophysical Research Communications

343

338

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The surface of endothelial cells is decorated with a wide variety of membrane-bound macromolecules that constitute the glycocalyx. These include glycoproteins bearing acidic oligosaccharides with terminal sialic acids (SA), and proteoglycans with their associated glycosaminoglycan that include: heparan sulfate (HS), chondroitin sulfate (CS) and hyaluronic acid (HA). In this study enzymes were used to selectively degrade glycoclyx components from the surface of bovine aortic endothelial cells and the effects of these alterations on fluid shear-induced nitric oxide (NO) and prostacyclin (PGI 2 ) production were determined. Depletion of HS, HA and SA, but not CS, blocked shear-induced NO production. Surprisingly, the same enzyme depletions that blocked NO production had no influence on shear-induced PGI 2 production. The results may be interpreted in terms of a glypicancaveolae-eNOS mechanism for shear-induced NO transduction, with PGI 2 being transduced in basal adhesion plaques that sense the same reaction stress whether the glycocalyx is intact or not.

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Section: Discussionmentioning

confidence: 99%

The role of endothelial glycocalyx components in mechanotransduction of fluid shear stress

Pahakis¹,

Kosky²,

Dull³

et al. 2007

Biochemical and Biophysical Research Communications

343

338

View full text Add to dashboard Cite

show abstract

“…The heterogeneous localization of eNOS is known to vary depending on the species, degree of confluency, passage, and even across the vascular tree (27,28,42,43). The mechanisms regulating the intracellular distribution of eNOS between the Golgi and plasma membrane pools and the physiological role underlying their distinct regulation remain to be established.…”

Section: Discussionmentioning

confidence: 99%

“…The majority of cellular eNOS and eNOS activity, normalized to caveola protein content, is concentrated in caveolin-1-enriched microdomains (25). Confluent endothelial cells have more plasma membrane and less Golgi eNOS than do subconfluent cells and release more NO in response to agonist challenge (42). The eNOS-binding proteins NOSIP and NOSTRIN displace plasma membrane eNOS to internal membranes (unlikely Golgi) and reduce calcium-dependent NO release (47,48).…”

Section: Discussionmentioning

confidence: 99%

“…It is well established that eNOS is localized to both the plasma membrane and the cytoplasmic aspects of the cis/medial-Golgi complex in endothelial cells both in intact blood vessels and in culture (25,27,28,30,33,(42)(43)(44)(45). More recently, transgenesis of the entire human eNOS gene encoding an eNOS-green fluorescent protein recapitulated imaging and functional studies of eNOS-green fluorescent protein in cultured cells (43) and unequivocally documented the bimodal distribution of eNOS into the Golgi complex and plasma membrane of endothelial cells lining intact blood vessels (46).…”

Section: Discussionmentioning

confidence: 99%

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Targeting of Endothelial Nitric-oxide Synthase to the Cytoplasmic Face of the Golgi Complex or Plasma Membrane Regulates Akt- Versus Calcium-dependent Mechanisms for Nitric Oxide Release

Fulton

Babbitt

Zoellner

et al. 2004

Journal of Biological Chemistry

126

125

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The heterogeneous localization of endothelial nitricoxide synthase (eNOS) on the Golgi complex versus the plasma membrane has made it difficult to dissect the regulation of each pool of enzyme. Here, we generated fusion proteins that specifically target the plasma membrane or cytoplasmic aspects of the Golgi complex and have assessed eNOS activation. Plasma membrane-targeted eNOS constructs were constitutively active, phosphorylated, and responsive to transmembrane calcium fluxes, yet were insensitive to further activation by Aktmediated phosphorylation. In contrast, cis-Golgi complex-targeted eNOS behaved similarly to wild-type eNOS and was less sensitive to calcium-dependent activation and highly responsive to Akt-dependent phosphorylation compared with plasma membrane versions. In plasma membrane-and Golgi complex-targeted constructs, Ser 1179 is critical for NO production. This study provides clear evidence for functional roles of plasma membrane-and Golgi complex-localized eNOS and supports the concept that proteins thought to be regulated and to function exclusively in the plasma membrane of cells can indeed signal and be regulated in internal Golgi membranes.Within the vascular endothelium, nitric oxide (NO) is generated via the enzyme endothelial nitric-oxide synthase (eNOS). 1

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“…eNOS, in an intracellular location-specific manner. Recent studies have demonstrated that cell-cell contact induces the enrichment of eNOS at intercellular junctions (Govers et al, 2002). Because eNOS is associated with PKA catalytic subunit at endothelial cell junctions (Heijnen et al, 2004), it would be interesting to examine whether their interaction is altered by shear stress.…”

Section: Shear Stress Stimulates Intracellular Translocation Of Pkamentioning

confidence: 99%

Shear stress stimulates phosphorylation of protein kinase A substrate proteins including endothelial nitric oxide synthase in endothelial cells

Boo

2006

Exp Mol Med

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Fluid shear stress plays a critical role in vascular health and disease. While protein kinase A (PKA) has been implicated in shear-stimulated signaling events in endothelial cells, it remains unclear whether and how PKA is stimulated in response to shear stress. This issue was addressed in the present study by monitoring the phosphorylation of endogenous substrates of PKA. Shear stress stimulated the phosphorylation of cAMP responsive element binding protein (CREB) in a PKA-dependent manner. Western blot analysis using the antibody reactive against the consensus motif of PKA substrates detected two proteins, P135 and P50, whose phosphorylation was increased by shear stress. The phosphorylation of P135 was blocked by a PKA inhibitor, H89, but not by a phosphoinositide 3-kinase inhibitor, wortmannin. Expression of a constitutively active PKA subunit stimulated P135 phosphorylation, supporting the potential of P135 as a PKA substrate. P135 was identified as endothelial nitric oxide synthase (eNOS) by immunoprecipitation study. PKA appeared to mediate shear stress-stimulated eNOS activation. Shear stress stimulated intracellular translocation of PKA activity from 'soluble' to 'particulate' fractions without involving cellular cAMP increase. Taken together, this study suggests that shear stress stimulates PKA-dependent phosphorylation of target proteins including eNOS, probably by enhancing intracellular site-specific interactions between protein kinase and substrates.

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Endothelial nitric oxide synthase activity is linked to its presence at cell–cell contacts

Cited by 68 publications

References 44 publications

The role of endothelial glycocalyx components in mechanotransduction of fluid shear stress

The role of endothelial glycocalyx components in mechanotransduction of fluid shear stress

Targeting of Endothelial Nitric-oxide Synthase to the Cytoplasmic Face of the Golgi Complex or Plasma Membrane Regulates Akt- Versus Calcium-dependent Mechanisms for Nitric Oxide Release

Shear stress stimulates phosphorylation of protein kinase A substrate proteins including endothelial nitric oxide synthase in endothelial cells

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