TNF-α-activated eNOS signaling increases leukocyte adhesion through the <i>S</i>- nitrosylation pathway

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dc.contributor.authorAguilar, Gaynor
dc.contributor.authorCordova, Francisco
dc.contributor.authorKoning, Tania
dc.contributor.authorSarmiento, Jose
dc.contributor.authorBoric, Mauricio P.
dc.contributor.authorBirukov, Konstantin
dc.contributor.authorCancino, Jorge
dc.contributor.authorVaras-Godoy, Manuel
dc.contributor.authorSoza, Andrea
dc.contributor.authorAlves, Natascha G.
dc.contributor.authorMujica, Patricio E.
dc.contributor.authorDuran, Walter N.
dc.contributor.authorEhrenfeld, Pamela
dc.contributor.authorSanchez, Fabiola A.
dc.date.accessioned2025-01-20T22:03:03Z
dc.date.available2025-01-20T22:03:03Z
dc.date.issued2021
dc.description.abstractNitric oxide ( NO) is a key factor in inflammation. Endothelial nitric oxide synthase (eNOS), whose activity increases after stimulation with proinflammatory cytokines, produces NO in endothelium. NO activates two pathways: 1) soluble guanylate cyclase-protein kinase G and 2) S-nitrosylation (NO-induced modification of free-thiol cysteines in proteins). S-nitrosylation affects phosphorylation, localization, and protein interactions. NO is classically described as a negative regulator of leukocyte adhesion to endothelial cells. However, agonists activating NO production induce a fast leukocyte adhesion, which suggests that NO might positively regulate leukocyte adhesion. We tested the hypothesis that eNOS-induced NO promotes leukocyte adhesion through the S-nitrosylation pathway. We stimulated leukocyte adhesion to endothelium in vitro and in vivo using tumor necrosis factor-alpha (TNF-alpha) as proinflammatory agonist. ICAM-1 changes were evaluated by immunofluorescence, subcellular fractionation, immunoprecipitation, and fluorescence recovery after photobleaching (FRAP). Protein kinase C sigma (PKC sigma) activity and S-nitrosylation were evaluated by Western blot analysis and biotin switch method, respectively. TNF-alpha, at short times of stimulation, activated the eNOS S-nitrosylation pathway and caused leukocyte adhesion to endothelial cells in vivo and in vitro. TNF-alpha-induced NO led to changes in ICAM-1 at the cell surface, which are characteristic of clustering. TNF-alpha-induced NO also produced S-nitrosylation and phosphorylation of PKCf, association of PKCf with ICAM-1, and ICAM-1 phosphorylation. The inhibition of PKCf blocked leukocyte adhesion induced by TNF-alpha. Mass spectrometry analysis of purified PKCf identified cysteine 503 as the only S-nitrosylated residue in the kinase domain of the protein. Our results reveal a new eNOS S-nitrosylation-dependent mechanism that induces leukocyte adhesion and suggests that S-nitrosylation of PKCf may be an important regulatory step in early leukocyte adhesion in inflammation.
dc.description.abstractNEW & NOTEWORTHY Contrary to the well-established inhibitory role of NO in leukocyte adhesion, we demonstrate a positive role of nitric oxide in this process. We demonstrate that NO induced by eNOS after TNF-alpha treatment induces early leukocyte adhesion activating the S-nitrosylation pathway. Our data suggest that PKCf S-nitrosylation may be a key step in this process.
dc.fuente.origenWOS
dc.identifier.doi10.1152/ajpheart.00065.2021
dc.identifier.eissn1522-1539
dc.identifier.issn0363-6135
dc.identifier.urihttps://doi.org/10.1152/ajpheart.00065.2021
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/94011
dc.identifier.wosidWOS:000744245000003
dc.issue.numero6
dc.language.isoen
dc.pagina.finalH1095
dc.pagina.inicioH1083
dc.revistaAmerican journal of physiology-heart and circulatory physiology
dc.rightsacceso restringido
dc.subjectleukocyte adhesion
dc.subjectnitric oxide
dc.subjectprotein kinase
dc.subjectS-nitrosylation
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleTNF-α-activated eNOS signaling increases leukocyte adhesion through the <i>S</i>- nitrosylation pathway
dc.typeartículo
dc.volumen321
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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