Phosphorylation of endothelial nitric oxide synthase by atypical PKCζ contributes to angiopoietin-1–dependent inhibition of VEGF-induced endothelial permeability in vitro

Oubaha, Malika; Gratton, Jean‐Philippe

doi:10.1182/blood-2008-12-196584

Cited by 59 publications

(50 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We demonstrate that PKC-z activation is necessary and sufficient to disrupt TJs and epithelial barrier in AECs, favoring the recruitment of neutrophils and the development of DEP-induced lung injury. PKC-z has been previously implicated in mediating alveolar epithelial disruption in response to hypoxia in alveolar epithelia (14) and the blood-brain barrier (13), as well as in the regulation of the endothelial barrier (32). This kinase also regulates occludin distribution and the assembly of TJs in Madin-Darby canine kidney and Caucasian colon adenocarcinoma (Caco-2) cells (33).…”

Section: Discussionmentioning

confidence: 99%

Protein Kinase C–ζ Mediates Lung Injury Induced by Diesel Exhaust Particles

Caraballo

Borcherding

Thorne

et al. 2013

Am J Respir Cell Mol Biol

View full text Add to dashboard Cite

Recently, we reported that diesel exhaust particles (DEPs) disrupt tight junctions (TJs) in alveolar epithelial cells (AECs) via an increase in reactive oxygen species (ROS). In this study, we investigated the role of protein kinase C (PKC)-z activation in DEP-induced lung injury. C57/bl6 mice were instilled intratracheally with 50 ml of saline containing 100 mg of DEPs or titanium dioxide (TiO 2 ). Twenty-four hours later, bronchoalveolar lavage was performed to assess neutrophil counts and protein concentrations. In addition, in vitro experiments were performed in primary rat and human AECs exposed to DEPs (50 mg/cm 2 ) for 3 hours. Transepithelial electrical conductance was measured, and TJ protein association was analyzed by immunoprecipitation. To determine whether the overexpression of antioxidants prevented DEP-induced lung injury, AECs and mice were infected with adenoviruses containing catalase and manganese superoxide dismutase (MnSOD) plasmids. In vivo, the overexpression of catalase and MnSOD prevented DEP-induced neutrophil recruitment. The inhibition of PKC-z activation also prevented DEP-induced neutrophil recruitment in vivo. In vitro, DEPs activated PKC-z in AECs, but not in alveolar macrophages. Using a specific myristolated PKC-z pseudosubstrate pepetide (PKC-z ps), we showed that PKC-z mediated the DEP-induced dissociation of occludin and zonula occludin-1 (ZO1) in rat and human AECs. In addition, the overexpression of constitutively active PKC-z induced the dissociation of occludin and ZO1 in AECs. DEP-induced TJ disruption occurs via PKC-z. TJ disruption seems to be in part responsible for DEP-induced lung injury.Keywords: diesel exhaust particles; PKC-z; occludin; ZO1; ROS Numerous pollutants exert detrimental effects on human health (1, 2). Diesel exhaust particles (DEPs), which are among the most abundant pollutants (3), comprise a component of particulate matter (PM) of less than 2.5 mm (PM 2.5 ) in size. The smallest particles are the most numerous, and can penetrate deepest into the lung, reaching the alveolar space (4). PM varies not only in size but in composition, which can include water-soluble and water-insoluble fine cores, with reactive and nonreactive surfaces. Soluble components of PM are the most bioavailable fraction of constituents playing a potential key role in the induction of adverse health effects, both at the site of their deposition in the lung, and in remote tissues and organ systems (5, 6). In addition, organic species can constitute a large portion of motor vehicle exhaust particles, and are considered among the most significant causative constituents of ambient PM. The short-term inhalation of DEPs produces systemic and pulmonary inflammation, enhances bronchial hyperresponsiveness in patients with asthma, and increases sensitization to airborne allergens (7-9). Furthermore, inhaling DEPs increases the number of inflammatory cells in the airway, and up-regulates inflammatory mediators (10).Lung injury is associated with a dysfunctional epithelial barrier allowin...

show abstract

Section: Discussionmentioning

confidence: 99%

Protein Kinase C–ζ Mediates Lung Injury Induced by Diesel Exhaust Particles

Caraballo

Borcherding

Thorne

et al. 2013

Am J Respir Cell Mol Biol

View full text Add to dashboard Cite

show abstract

“…12 To identify molecular players linking PKC signaling events downstream of the Ang-1 receptor Tie2, we used immunoprecipitation coupled to mass spectrometry and identified ␤-catenin as a PKC interacting protein in lysates from Ang-1-stimulated endothelial cells (supplemental Table 1). We show that, simultaneous to the phosphorylation of PKC, Ang-1 stimulation induced the association of ␤-catenin with PKC ( Figure 2A).…”

Section: Association Of Pkc With ␤-Catenin In Endothelial Cellsmentioning

confidence: 99%

“…However, it is well known that they induce opposite effects on adherens junctions and on endothelial cell permeability. [10][11][12] VEGF promotes the dissociation of adherens junctions, whereas Ang-1 induces the stabilization of junctional complexes. [12][13][14] Interestingly, signaling at cell-cell junctions can serve as polarity cues to organize cell movement and general organization of cell shape.…”

Section: Introductionmentioning

confidence: 99%

“…[10][11][12] VEGF promotes the dissociation of adherens junctions, whereas Ang-1 induces the stabilization of junctional complexes. [12][13][14] Interestingly, signaling at cell-cell junctions can serve as polarity cues to organize cell movement and general organization of cell shape. 15 In migrating mammalian cells, Par/atypical protein kinase C (PKC) polarity complexes are recruited to the leading edge and have been implicated in the initial polarization events necessary for directed cell migration of astrocytes and epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Formation of a PKCζ/β-catenin complex in endothelial cells promotes angiopoietin-1–induced collective directional migration and angiogenic sprouting

Oubaha

Lin

Margaron

et al. 2012

Blood

Self Cite

View full text Add to dashboard Cite

“…209,210 Only one paper has noted the presence of any PKC isoform at the MEJ, and PKC itself was not the focus of that paper. 12 There is precedent for PKC regulation of eNOS phosphorylation [211][212][213][214] and the phosphorylation of eNOS being dependent on ER-derived calcium. 215 The MEJ lipidomic results presented here point to, for the first time, 1) an ideal lipid environment for PKC based on the enrichment of PS and DAG and 2) a unique lipid environment in this arterial model as compared to EC and SMC grown in alone and in non-polarized conditions.…”

Section: Discussionmentioning

confidence: 99%

Signaling Microdomains within the Myoendothelial Junction

Biwer

View full text Add to dashboard Cite

Communication between endothelial and smooth muscle cells is critical to maintain homeostatic vascular tone and blood pressure. In resistance arteries, the cells make direct cytoplasmic contact via cellular extensions that project through the thin extracellular matrix of the internal elastic lamina that separates them. These structures are termed myoendothelial junctions (MEJ). There are a number of proteins and signaling processes localized to this part of the arterial wall that regulate bi-directional communication between the endothelium and smooth muscle. In particular, a number of proteins localized to the MEJ mediate calcium signaling and there is localized calcium release from the endoplasmic reticulum via the inositol (1, 4, 5) trisphosphate receptor that can influence vascular tone and negative feedback. There is a detailed description of the creation and isolation of in vitro MEJ using the vascular cell co-culture technique, which has allowed for deeper exploration of signaling molecules and processes occurring at the MEJ. Smooth muscle cells and endothelial cells grow on opposite sides of a filter inset, and the MEJ form within the holes of the filter. This model can be used to immunofluorescently label proteins or the cellular fractions can be individually isolated to investigate protein expression and phosphorylation via western blotting. Using this unique model, we show the in vitro MEJ has a unique lipid composition that is rich in diacylglycerol and phosphatidylserine. This likely facilitates localization of signaling molecules such as protein kinase C and selective activation of endothelial nitric oxide synthase. Additionally, the multifunctional protein calreticulin is highly expressed at the MEJ of resistance arteries and endothelial deletion of calreticulin results in impaired MEJ calcium signaling, thus vasoconstriction to phenylephrine and blood pressure. Our data suggests that EC monolayer Calr could be iii functionally different than MEJ-localized Calr, which may be located outside the ER.Further work will need to be done in order to clarify and confirm MEJ Calr function and how it affects calcium signaling at the MEJ specifically in response to phenylephrine. In summary, the MEJ is an important signaling microdomain strategically located in the wall of resistance arteries to mediate heterocellular communication.

show abstract

Phosphorylation of endothelial nitric oxide synthase by atypical PKCζ contributes to angiopoietin-1–dependent inhibition of VEGF-induced endothelial permeability in vitro

Cited by 59 publications

References 48 publications

Protein Kinase C–ζ Mediates Lung Injury Induced by Diesel Exhaust Particles

Protein Kinase C–ζ Mediates Lung Injury Induced by Diesel Exhaust Particles

Formation of a PKCζ/β-catenin complex in endothelial cells promotes angiopoietin-1–induced collective directional migration and angiogenic sprouting

Signaling Microdomains within the Myoendothelial Junction

Contact Info

Product

Resources

About