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• W4310731609 abstract "Hyperlipidemia characterized by high blood levels of free fatty acids (FFAs) is important for the progression of inflammatory cardiovascular diseases. Integrin β1 is a transmembrane receptor that drives various cellular functions, including differentiation, migration, and phagocytosis. However, the underlying mechanisms modifying integrin β1 protein and activity in mediating monocyte/macrophage adhesion to endothelium remain poorly understood. In this study, we demonstrated that integrin β1 protein underwent S-nitrosylation in response to nitrosative stress in macrophages. To examine the effect of elevated levels of FFA on the modulation of integrin β1 expression, we treated the macrophages with a combination of oleic acid and palmitic acid (2:1) and found that FFA activated inducible nitric oxide synthase/nitric oxide and increased the integrin β1 protein level without altering the mRNA level. FFA promoted integrin β1 S-nitrosylation via inducible nitric oxide synthase/nitric oxide and prevented its degradation by decreasing binding to E3 ubiquitin ligase c-Cbl. Furthermore, we found that increased integrin α4β1 heterodimerization resulted in monocyte/macrophage adhesion to endothelium. In conclusion, these results provided novel evidence that FFA-stimulated N--O stabilizes integrin β1 via S-nitrosylation, favoring integrin α4β1 ligation to promote vascular inflammation. Hyperlipidemia characterized by high blood levels of free fatty acids (FFAs) is important for the progression of inflammatory cardiovascular diseases. Integrin β1 is a transmembrane receptor that drives various cellular functions, including differentiation, migration, and phagocytosis. However, the underlying mechanisms modifying integrin β1 protein and activity in mediating monocyte/macrophage adhesion to endothelium remain poorly understood. In this study, we demonstrated that integrin β1 protein underwent S-nitrosylation in response to nitrosative stress in macrophages. To examine the effect of elevated levels of FFA on the modulation of integrin β1 expression, we treated the macrophages with a combination of oleic acid and palmitic acid (2:1) and found that FFA activated inducible nitric oxide synthase/nitric oxide and increased the integrin β1 protein level without altering the mRNA level. FFA promoted integrin β1 S-nitrosylation via inducible nitric oxide synthase/nitric oxide and prevented its degradation by decreasing binding to E3 ubiquitin ligase c-Cbl. Furthermore, we found that increased integrin α4β1 heterodimerization resulted in monocyte/macrophage adhesion to endothelium. In conclusion, these results provided novel evidence that FFA-stimulated N--O stabilizes integrin β1 via S-nitrosylation, favoring integrin α4β1 ligation to promote vascular inflammation. Hyperlipidemia manifested by high blood levels of free fatty acids (FFAs) is critical for the progression of multiple metabolic diseases as well as cardiovascular complications. FFA levels increase significantly in obesity, type 2 diabetes, and atherosclerosis (1Boden G. Obesity and free fatty acids.Endocrinol. Metab. Clin. North Am. 2008; 37 (viii-ix): 635-646Abstract Full Text Full Text PDF PubMed Scopus (577) Google Scholar, 2Prescott J. Owens D. Collins P. Johnson A. Tomkin G.H. The fatty acid distribution in low density lipoprotein in diabetes.Biochim. Biophys. Acta. 1999; 1439: 110-116Crossref PubMed Scopus (21) Google Scholar, 3Rinne P. Guillamat-Prats R. Rami M. Bindila L. Ring L. Lyytikainen L.P. et al.Palmitoylethanolamide promotes a proresolving macrophage phenotype and attenuates atherosclerotic plaque formation.Arterioscler Thromb. Vasc. Biol. 2018; 38: 2562-2575Crossref PubMed Scopus (49) Google Scholar). Increased FFA level causes lipotoxicity and activates the release of proinflammatory cytokines such as IL-6, TNFα, and other endogenous endotoxin. These events are not only thought to link metabolic inflammation but also are even considered as cause of atherosclerosis and hypertension. In response to increased lipid levels, the endothelial cells release chemokines, whereas the monocyte/macrophages develop oxidative stress and upregulate chemokine receptor. Inducible nitric oxide synthase (iNOS) is the type of NOS expressed in monocyte/macrophage and plays a critical role in physiopathological processes ranging from antimicrobial and wound healing to proinflammatory diseases. Expression of iNOS is negligible in resting cells and induced upon infection or metabolic stress, such as elevated levels of glucose and lipid (4Anavi S. Tirosh O. iNOS as a metabolic enzyme under stress conditions.Free Radic. Biol. Med. 2020; 146: 16-35Crossref PubMed Scopus (91) Google Scholar). Accumulating evidence strongly implies that iNOS-derived nitric oxide (NO) plays a central role in the regulation of lipid metabolism during inflammatory conditions (5Rosas-Ballina M. Guan X.L. Schmidt A. Bumann D. Classical activation of macrophages leads to lipid droplet formation without de novo fatty acid synthesis.Front Immunol. 2020; 11: 131Crossref PubMed Scopus (30) Google Scholar), resulting in endothelial dysfunction, atherosclerosis, or hypertension (6Gliozzi M. Scicchitano M. Bosco F. Musolino V. Carresi C. Scarano F. et al.Modulation of nitric oxide synthases by oxidized LDLs: role in vascular inflammation and atherosclerosis development.Int. J. Mol. Sci. 2019; 20: 3294Crossref PubMed Scopus (93) Google Scholar, 7Fysikopoulos A. Seimetz M. Hadzic S. Knoepp F. Wu C.Y. Malkmus K. et al.Amelioration of elastase-induced lung emphysema and reversal of pulmonary hypertension by pharmacological iNOS inhibition in mice.Br. J. Pharmacol. 2021; 178: 152-171Crossref PubMed Scopus (10) Google Scholar, 8Cromheeke K.M. Kockx M.M. De Meyer G.R. Bosmans J.M. Bult H. Beelaerts W.J. et al.Inducible nitric oxide synthase colocalizes with signs of lipid oxidation/peroxidation in human atherosclerotic plaques.Cardiovasc. Res. 1999; 43: 744-754Crossref PubMed Scopus (107) Google Scholar). Adhesive interactions between immune and other cell types are largely mediated by integrins. Mammalian integrins comprise 18 α and eight β subunits that constitute 24 heterodimers, which have been implicated in either acute (9Mezu-Ndubuisi O.J. Maheshwari A. The role of integrins in inflammation and angiogenesis.Pediatr. Res. 2021; 89: 1619-1626Crossref PubMed Scopus (97) Google Scholar) or chronic metabolic inflammation (10Tian Y. Yang C. Yao Q. Qian L. Liu J. Xie X. et al.Procyanidin B2 activates PPARgamma to induce M2 polarization in mouse macrophages.Front Immunol. 2019; 10: 1895Crossref PubMed Scopus (38) Google Scholar). Integrin β1 serves a critical function in cell adhesion and occurs on numerous cells of the immune system (11Mrugacz M. Bryl A. Falkowski M. Zorena K. Integrins: an important link between angiogenesis, inflammation and eye diseases.Cells. 2021; 10: 1703Crossref PubMed Scopus (17) Google Scholar). The evidence that integrin α4β1 heterodimer modulates metabolic inflammation in obesity implies its role in leukocyte recruitment (12Chung K.J. Chatzigeorgiou A. Economopoulou M. Garcia-Martin R. Alexaki V.I. Mitroulis I. et al.A self-sustained loop of inflammation-driven inhibition of beige adipogenesis in obesity.Nat. Immunol. 2017; 18: 654-664Crossref PubMed Scopus (111) Google Scholar). Integrin α4β1, presented on the leukocyte membrane, binds VCAM-1 on endothelial cells (13Gorina R. Lyck R. Vestweber D. Engelhardt B. Beta2 integrin-mediated crawling on endothelial ICAM-1 and ICAM-2 is a prerequisite for transcellular neutrophil diapedesis across the inflamed blood-brain barrier.J. Immunol. 2014; 192: 324-337Crossref PubMed Scopus (111) Google Scholar, 14Chang A.C. Chen P.C. Lin Y.F. Su C.M. Liu J.F. Lin T.H. et al.Osteoblast-secreted WISP-1 promotes adherence of prostate cancer cells to bone via the VCAM-1/integrin alpha4beta1 system.Cancer Lett. 2018; 426: 47-56Crossref PubMed Scopus (44) Google Scholar). Rapid chemokine-increased integrin α4β1 affinity is required for monocyte arrest (15Hyduk S.J. Rullo J. Cano A.P. Xiao H. Chen M. Moser M. et al.Talin-1 and kindlin-3 regulate alpha4beta1 integrin-mediated adhesion stabilization, but not G protein-coupled receptor-induced affinity upregulation.J. Immunol. 2011; 187: 4360-4368Crossref PubMed Scopus (33) Google Scholar). During inflammation, integrin α4β1 drives transendothelial leukocyte migration into the inflamed tissue (16Sackstein R. The lymphocyte homing receptors: gatekeepers of the multistep paradigm.Curr. Opin. Hematol. 2005; 12: 444-450Crossref PubMed Scopus (110) Google Scholar). In addition, integrin α4β1 can interact with vascular endothelial growth factor receptor 2, contributing to vascular endothelial growth factor functions (17Gutierrez-Gonzalez A. Aguilera-Montilla N. Ugarte-Berzal E. Bailon E. Cerro-Pardo I. Sanchez-Maroto C. et al.Alpha4beta1 integrin associates with VEGFR2 in CLL cells and contributes to VEGF binding and intracellular signaling.Blood Adv. 2019; 3: 2144-2148Crossref PubMed Scopus (7) Google Scholar). Integrin β1 expression is regulated during diverse pathophysiological processes including infection, tumorigenesis, and asthma (18Wang Y. Li K. Zhao W. Liu Z. Liu J. Shi A. et al.Aldehyde dehydrogenase 3B2 promotes the proliferation and invasion of cholangiocarcinoma by increasing integrin beta 1 expression.Cell Death Dis. 2021; 12: 1158Crossref PubMed Scopus (8) Google Scholar, 19Teoh C.M. Tan S.S. Tran T. Integrins as therapeutic targets for respiratory diseases.Curr. Mol. Med. 2015; 15: 714-734Crossref PubMed Scopus (38) Google Scholar, 20Speziale P. Pietrocola G. The multivalent role of fibronectin-binding proteins A and B (FnBPA and FnBPB) of staphylococcus aureus in host infections.Front Microbiol. 2020; 11: 2054Crossref PubMed Scopus (36) Google Scholar). A change in the expression of integrin receptors usually takes place at the transcription level. However, integrin β1 protein undergoes posttranslational modifications, including phosphorylation (21Gahmberg C.G. Gronholm M. How integrin phosphorylations regulate cell adhesion and signaling.Trends Biochem. Sci. 2022; 47: 265-278Abstract Full Text Full Text PDF PubMed Scopus (17) Google Scholar), acetylation (22Mirgorodskaya E. Dransart E. Shafaq-Zadah M. Roderer D. Sihlbom C. Leffler H. et al.Site-specific N-glycan profiles of alpha5 beta1 integrin from rat liver.Biol. Cell. 2022; 114: 160-176Crossref PubMed Scopus (1) Google Scholar) and ubiquitination (23Fan L. Zhang Y. Shi D. Xi R. Zhang Z. Wang X. Hypoxia enhances the cytotoxic effect of As4S4 on rat ventricular H9c2 cells through activation of ubiquitin-proteasome system.J. Trace Elem. Med. Biol. 2021; 66: 126720Crossref PubMed Scopus (2) Google Scholar). Our understanding of the other posttranslational modification of integrin β1 is limited. Protein S-nitrosylation is a redox sensitive reaction at cysteine residues by NO or NO-derived species to generate a S-nitrosothiol (24Chatterji A. Banerjee D. Billiar T.R. Sengupta R. Understanding the role of S-nitrosylation/nitrosative stress in inflammation and the role of cellular denitrosylases in inflammation modulation: implications in health and diseases.Free Radic. Biol. Med. 2021; 172: 604-621Crossref PubMed Scopus (5) Google Scholar). Previous reports suggest that certain posttranslational modification may regulate protein ubiquitination and function. S-nitrosylation may promote or inhibit degradation of certain protein such as dihydrofolate reductase (25Cai Z. Lu Q. Ding Y. Wang Q. Xiao L. Song P. et al.Endothelial nitric oxide synthase-derived nitric oxide prevents dihydrofolate reductase degradation via promoting S-nitrosylation.Arterioscler Thromb. Vasc. Biol. 2015; 35: 2366-2373Crossref PubMed Scopus (21) Google Scholar) or TRIM72 (26Kohr M.J. Evangelista A.M. Ferlito M. Steenbergen C. Murphy E. S-nitrosylation of TRIM72 at cysteine 144 is critical for protection against oxidation-induced protein degradation and cell death.J. Mol. Cell Cardiol. 2014; 69: 67-74Abstract Full Text Full Text PDF PubMed Scopus (53) Google Scholar), respectively. S-nitrosylation of integrin α6 decreases its binding to laminin-β1 and increases the extent of prostate cancer cell migration (27Isaac J. Tarapore P. Zhang X. Lam Y.W. Ho S.M. Site-specific S-nitrosylation of integrin alpha6 increases the extent of prostate cancer cell migration by enhancing integrin beta1 association and weakening adherence to laminin-1.Biochemistry. 2012; 51: 9689-9697Crossref PubMed Scopus (18) Google Scholar). Thus, whether integrin β1 could be regulated through S-nitrosylation remains as a critical question to understand its signaling transduction and function. In this study, we demonstrated that iNOS-generated NO stabilizes integrin β1 via S-nitrosylation, favorizing integrin α4β1 ligation and monocyte/macrophage adhesion to endothelium. Expression of iNOS is upregulated upon metabolic perturbation and considered as a marker of the classical and proinflammatory activation of macrophages (4Anavi S. Tirosh O. iNOS as a metabolic enzyme under stress conditions.Free Radic. Biol. Med. 2020; 146: 16-35Crossref PubMed Scopus (91) Google Scholar). To investigate whether high level of FFA could induce inflammation in macrophages, Raw264.7 cells were subjected to FFA for the indicated times. FFA increased the levels of iNOS mRNA (Fig. 1A) and protein (Fig. 1B) 4 h after treatment. Then, the effect of FFA on NO production was examined in macrophages. As shown in Fig. 1C, treatment with FFA significantly augmented NO quantity in the conditioned media of macrophages. This result was further verified using an electron paramagnetic resonance (EPR) spectroscopy (Fig. 1D). These data demonstrated that iNOS expression was upregulated by FFA in macrophages. iNOS plays a critical role in inflammatory progression, promoting us to evaluate its precise regulation on integrin β1. Raw264.7 cells were transfected with scrambled siRNA or two pairs of iNOS-specific siRNA. Both iNOS siRNAs markedly reduced iNOS protein compared to those transfected with scrambled siRNA, indicating their specificity. iNOS silencing reduced FFA-increased integrin β1 protein level compared to that of macrophages transfected with scrambled siRNA (Fig. 2A). However, the integrin β1 mRNA level was not altered (Fig. 2B). Similar to the results of iNOS siRNA silencing, 1400W, a selective iNOS inhibitor, ablated FFA-induced integrin β1 expression (Fig. 2C). These data suggested that iNOS participated in FFA-increased integrin β1 protein level in macrophages. Next, we investigated the effect of iNOS-derived NO on integrin β1 expression. Raw264.7 cells were exposed to 2-Phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), a known NO scavenger. Treatment of macrophages with PTIO (60 μM and 120 μM) for 24 h abolished FFA-induced integrin β1 protein level (Fig. 2D) without affecting iNOS expression (Fig. S1). Exposure of macrophages to PTIO (60 μM) for 24 h significantly reduced FFA-induced integrin β1 (Fig. 2E). These data suggested that FFA-induced NO augmented integrin β1 protein level. Since both iNOS silencing and NO scavenging lowered integrin β1 protein without altering its transcription, we reasoned whether NO would modulate integrin β1 protein. As depicted in Fig. 3A, exposure of Raw264.7 cells to S-nitrosoglutathion (GSNO) or organic salt sodium nitroprusside (SNP), two NO donors, augmented integrin β1 protein level, which was abolished by a reductant N-acetyl-L-cysteinamide (NAC). Taken together, these results suggested that NO might modulate integrin β1 protein level via a redox-sensitive reaction, promoting us to examine whether integrin β1 could be modified by S- nitrosylation. Raw264.7 cells were exposed to GSNO or SNP for 4 h and then subjected to irreversible biotinylation procedure. Immunoblotting showed marked S-nitrosylation of integrin β1, which was abrogated by NAC treatment (Fig. 3B). These data suggested that integrin β1 could be S-nitrosylated. Experiments conducted in murine bone marrow–derived macrophages also confirmed that NO might modulate integrin β1 protein level (Fig. 3C) via a S-nitrosylation-dependent mechanism (Fig. 3D). Integrin β1 protein contains cysteine-rich tandem repeats which are crucial for its activation (28Eble J.A. de Rezende F.F. Redox-relevant aspects of the extracellular matrix and its cellular contacts via integrins.Antioxid. Redox Signal. 2014; 20: 1977-1993Crossref PubMed Scopus (65) Google Scholar). As shown in Figure 3E, three cysteine residues (Cys536, Cys555, and Cys568), highly conserved among species, were predicted as potential S-nitrosylated sites by using three different web-based tools (iSNO-PseACC, iSNO-AAPair, and PreSNO). To further confirm the function residue that undergo S-nitrosylation, we replaced the three corresponding cysteines in integrin β1 with alanine (C536A, C555A, and C568A) by using site-directed mutagenesis. The expression plasmids of the wildtype or mutated integrin β1 was transfected into HEK 293 cells and then treated with GSNO. We found that integrin β1 mutation at C555A, but not C536A or C568A abolished GNSO-induced S-nitrosylation (Fig. 3F), suggesting that S-nitrosylation modified integrin β1 predominantly at Cys555. Because iNOS inhibitor attenuated the FFA-stimulated increase in integrin β1 protein level without affecting its mRNA level (Fig. 2B), we studied the effect of NO on integrin β1 protein stability. HEK 293 cells were transfected with plasmid expressing integrin β1. Then the cells were exposed to GSNO after pretreated with cycloheximide (CHX), an inhibitor of de novo protein synthesis. As shown in Figure 4A, GSNO increased the half-life of integrin β1 protein. It is reported that integrin β1 degradation was mediated by E3 ubiquitin ligase c-Cbl (23Fan L. Zhang Y. Shi D. Xi R. Zhang Z. Wang X. Hypoxia enhances the cytotoxic effect of As4S4 on rat ventricular H9c2 cells through activation of ubiquitin-proteasome system.J. Trace Elem. Med. Biol. 2021; 66: 126720Crossref PubMed Scopus (2) Google Scholar). Co-immunoprecipitation indicated that the binding of c-Cbl to integrin β1 was largely diminished by GSNO treatment (Fig. 4B), which is consistent with the result of ubiquitination. As depicted in Figure 4C, GSNO decreased the detection of ubiquitinated integrin β1, suggesting that NO maintained integrin β1 stability by decreasing c-Cbl-dependent ubiquitination. As increasement of integrin β1 protein level was dependent on iNOS-derived NO, we reasoned whether integrin β1 would be modified by S-nitrosylation. For this purpose, Raw264.7 and bone marrow–derived macrophages were exposed to FFA. Immunoblotting showed significant S-nitrosylation of integrin β1, which was abolished by NAC pretreatment (Fig. 5A). Similar with the result of GSNO, macrophages with FFA treatment after 24 h showed an increasement of integrin β1 protein level, which was ablated by NAC (Fig. 5B). These data suggested FFA could increase integrin β1 protein level via S-nitrosylation-dependent mechanism. We next examined whether iNOS silencing leads to integrin β1 reduction via ubiquitin-proteasome degradation. As shown in Figure 5C, MG132 ablated integrin β1 reduction caused by iNOS silencing. Similar with the results of GSNO, FFA treatment significantly inhibited integrin β1 degradation in the presence of CHX (Fig. 5D). Co-immunoprecipitation indicated that the binding of E3 ubiquitin ligase c-Cbl to integrin β1 was significantly decreased by FFA treatment (Fig. 5E), which was consistent that FFA decreased the ubiquitination level of integrin β1 (Fig. 5F). To confirm the effect of c-Cbl on integrin β1 degradation, Raw264.7 cells were primarily transfected with scrambled or c-Cbl siRNA, followed with FFA treatment. As depicted in Figure 5G, c-Cbl silencing further stabilized integrin β1 protein in FFA-treated macrophages. These data indicated that FFA augmented integrin β1 stability by reducing c-Cbl mediated ubiquitination. Given that integrins are obligate heterodimers and integrin α4 binds to integrin β1 in monocyte/macrophages (29Hyduk S.J. Chan J.R. Duffy S.T. Chen M. Peterson M.D. Waddell T.K. et al.Phospholipase C, calcium, and calmodulin are critical for alpha4beta1 integrin affinity up-regulation and monocyte arrest triggered by chemoattractants.Blood. 2007; 109: 176-184Crossref PubMed Scopus (81) Google Scholar), we examined the modulation of integrin α4 and β1 by FFA. FFA did not alter their mRNA expression (Fig. 6, A and B). Protein level of integrin β1 was induced by FFA in a concentration- (Fig. 6C) and time-dependent manner (Fig. 6D). In contrast, the protein level of integrin α4 remained unchanged (Fig. 6, C and D). Immunoprecipitation analysis was performed to examine the effect of FFA on integrin a4β1 dimerization. Their ligation was significantly enhanced in macrophages by FFA (Fig. 6E). Immunofluorescence staining on Raw274.7 cells confirmed that FFA upregulated integrin β1 protein level. Integrin α4 and β1 were localized on the surface of FFA-treated cells (Fig. 6F), suggesting that the formation of integrin α4β1 heterodimer on cellular membrane was increased by FFA in macrophages. To investigate the function and related mechanism of integrin β1 in macrophages, FFA-treated Raw264.7 cells were primarily transfected with integrin β1 or iNOS siRNA. Either integrin β1 (Fig. 7A) or iNOS (Fig. 7B) siRNA markedly reduced the number of adhering macrophages to endothelial cells compared to those transfected with scrambled siRNA. Similar to the effect of iNOS siRNA, selective iNOS inhibitor, 1400W, abolished the stimulatory effect of FFA on monocyte/macrophage-endothelial interaction (Fig. 7C). In parallel, NAC pretreatment decreased FFA-induced monocyte/macrophage adhesion (Fig. 7D). These results indicated that integrin β1 participated in FFA-induced monocyte/macrophage to endothelium via iNOS/NO-dependent S-nitrosylation. In this study, we revealed a novel modification of integrin β1, which is a key integrin subunit with essential roles in leukocyte extravascular migration and inflammatory responses. Here we demonstrated for the first time that elevated FFA triggered S-nitrosylation of integrin β1 in monocytes/macrophages. The decreased ubiquitination and proteasome-dependent degradation of integrin β1 enhanced monocyte–endothelial adhesion. Furthermore, these results provide mechanistic evidence that S-nitrosylation regulated the formation of a specific integrin heterodimer to mediate monocyte/macrophage adhesion to vascular endothelium. Integrins are essential for the survival and migratory capacity of inflammatory cells. Integrin β1 upregulation is a major mechanism for monocyte/macrophage adhesion, whereas its downregulation is crucial for cell migration and metastasis (30Shi L. Liu B. Shen D.D. Yan P. Zhang Y. Tian Y. et al.A tumor-suppressive circular RNA mediates uncanonical integrin degradation by the proteasome in liver cancer.Sci. Adv. 2021; 7eabe5043Crossref Scopus (29) Google Scholar). Therefore, integrin β1 serves as a switch to control cellular motility and recruitment. Integrin β1+ macrophages are highly detectable in carotid plaques of the patients with hyperlipidemia or in atherosclerotic lesions of ApoE−/− mice (31Huang L.H. Melton E.M. Li H. Sohn P. Rogers M.A. Mulligan-Kehoe M.J. et al.Myeloid Acyl-CoA:cholesterol Acyltransferase 1 deficiency reduces lesion macrophage content and suppresses atherosclerosis progression.J. Biol. Chem. 2016; 291: 6232-6244Abstract Full Text Full Text PDF PubMed Scopus (29) Google Scholar, 32Riek A.E. Oh J. Sprague J.E. Timpson A. de las Fuentes L. Bernal-Mizrachi L. et al.Vitamin D suppression of endoplasmic reticulum stress promotes an antiatherogenic monocyte/macrophage phenotype in type 2 diabetic patients.J. Biol. Chem. 2012; 287: 38482-38494Abstract Full Text Full Text PDF PubMed Scopus (88) Google Scholar). Oxysterol, a cholesterol oxidation product, causes strong expression of integrin β1 in human macrophage lineage via c-Src/PLC/PKC/ERK pathway (33Gargiulo S. Gamba P. Testa G. Sottero B. Maina M. Guina T. et al.Molecular signaling involved in oxysterol-induced beta(1)-integrin over-expression in human macrophages.Int. J. Mol. Sci. 2012; 13: 14278-14293Crossref PubMed Scopus (12) Google Scholar). Emerging evidence suggest that iNOS plays a critical role in metainflammation by creating nitrosative stress (34Bruno A.S. Lopes P.D.D. de Oliveira K.C.M. de Oliveira A.K. de Assis Cau S.B. Vascular inflammation in hypertension: targeting lipid mediators unbalance and nitrosative stress.Curr. Hypertens. Rev. 2021; 17: 35-46Crossref PubMed Scopus (5) Google Scholar). S-nitrosylation of integrin α6 at Cys86 decreases its binding to laminin-β1, leading to decreased cell adherence and increased motility in prostate cancer cells (27Isaac J. Tarapore P. Zhang X. Lam Y.W. Ho S.M. Site-specific S-nitrosylation of integrin alpha6 increases the extent of prostate cancer cell migration by enhancing integrin beta1 association and weakening adherence to laminin-1.Biochemistry. 2012; 51: 9689-9697Crossref PubMed Scopus (18) Google Scholar). Exogenous NO modulates integrin αIIbβ3 activation via a S-nitrosylation-dependent mechanism (35Walsh G.M. Leane D. Moran N. Keyes T.E. Forster R.J. Kenny D. et al.S-Nitrosylation of platelet alphaIIbbeta3 as revealed by Raman spectroscopy.Biochemistry. 2007; 46: 6429-6436Crossref PubMed Scopus (48) Google Scholar). However, the mechanism by which integrin undergoing S-nitrosylation is largely unknown. Our current study provided a mechanistic explanation as to how integrin β1 promoted monocyte-endothelial adhesion via S-nitrosylation, ubiquitination, and degradation. This finding may shed new light on the mechanisms by which dyslipidemia-associated metabolic perturbation leads to meta-inflammation. Several studies suggest that posttranslational modifications are required for integrin β1 signaling transduction. Glycosylation protein controls integrin β1 activity and endocytic trafficking (22Mirgorodskaya E. Dransart E. Shafaq-Zadah M. Roderer D. Sihlbom C. Leffler H. et al.Site-specific N-glycan profiles of alpha5 beta1 integrin from rat liver.Biol. Cell. 2022; 114: 160-176Crossref PubMed Scopus (1) Google Scholar). Phosphorylation of tyrosine 788 in integrin β1 promotes inside-out receptor activation (36Nilsson S. Kaniowska D. Brakebusch C. Fassler R. Johansson S. Threonine 788 in integrin subunit beta1 regulates integrin activation.Exp. Cell Res. 2006; 312: 844-853Crossref PubMed Scopus (24) Google Scholar). Here, we found that S-nitrosylation inhibited integrin β1 degradation and triggers monocyte/macrophage adhesion to endothelium. It was reported in a redox proteome research that the Cys568 might be another potential cysteine residue undergoing S-nitrosylation in N-acetyl-p-aminophenol treated-hepatocytes (37Wojdyla K. Wrzesinski K. Williamson J. Fey S.J. Rogowska-Wrzesinska A. Acetaminophen-induced S-nitrosylation and S-sulfenylation signalling in 3D cultured hepatocarcinoma cell spheroids.Toxicol. Res. (Camb). 2016; 5: 905-920Crossref PubMed Google Scholar). Both Cys555 and Cys568 are conserved among mouse, human, and rat. They are contained within the cysteine-rich tandem repeats which form disulfide bridge. The disulfide bridge is required for recognition of α subunit and critical for the transition from its inactive to active form (28Eble J.A. de Rezende F.F. Redox-relevant aspects of the extracellular matrix and its cellular contacts via integrins.Antioxid. Redox Signal. 2014; 20: 1977-1993Crossref PubMed Scopus (65) Google Scholar). This is consistent with our finding that S-nitrosylation of integrin β1 regulated its ligation with the α4 subunit. S-nitrosylation is reported to regulate protein ubiquitination and stability. Our research showed that S-nitrosylation decreased the binding between integrin β1 and E3 ubiquitin ligase c-Cbl, thereby inhibiting its ubiquitination and degradation. The mechanism by which S-nitrosylation prevents the interaction between integrin β1 and c-Cbl is to be investigated. A conformational change of integrin β1 might prevent recognition and subsequent attachment of ubiquitin. Other E3 ligases, such as Fbx2, are also involved in integrin β1 ubiquitination and proteasome-dependent degradation (30Shi L. Liu B. Shen D.D. Yan P. Zhang Y. Tian Y. et al.A tumor-suppressive circular RNA mediates uncanonical integrin degradation by the proteasome in liver cancer.Sci. Adv. 2021; 7eabe5043Crossref Scopus (29) Google Scholar). Integrin β1 has been shown to be ubiquitinated, internalized, and degraded by the lysosome, a typical degradation pathway for transmembrane proteins (38Bottcher R.T. Stremmel C. Meves A. Meyer H. Widmaier M. Tseng H.Y. et al.Sorting nexin 17 prevents lysosomal degradation of beta1 integrins by binding to the beta1-integrin tail.Nat. Cell Biol. 2012; 14: 584-592Crossref PubMed Scopus (154) Google Scholar). However, we found in this study that iNOS-derived NO prevented integrin β1 from proteasome-dependent degradation. It is tempting to speculate that the S-nitrosylation may mediate an unusual process for substrate recognition by the proteasome. Integrin are heterodimers of α and β subunits, which are strictly ligated upon extracellular stimuli and required for its downstream signaling activation. The mechanism of integrin dimerization has not been well-understood. C1q-containing immune complexes is required for integrin α2β1 ligation in mast cells during innate immunity (39McCall-Culbreath K.D. Li Z. Zutter M.M. Crosstalk between the alpha2beta1 integrin and c-met/HGF-R regulates innate immunity.Blood. 2008; 111: 3562-3570Crossref PubMed Scopus (45) Google Scholar). Peroxisome proliferator-activated receptor γ inhibits integrin αMβ2 and favorizes integrin αVβ5 ligation during macrophage M2 polarization (40Yao Q. Liu J. Zhang Z. Li F. Zhang C. Lai B. et" @default.
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• W4310731609 date "2023-01-01" @default.
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• W4310731609 title "Free fatty acids stabilize integrin β1 via S-nitrosylation to promote monocyte–endothelial adhesion" @default.
• W4310731609 cites W1896805280 @default.
• W4310731609 cites W1969156397 @default.
• W4310731609 cites W1979721564 @default.
• W4310731609 cites W1992283755 @default.
• W4310731609 cites W2001422785 @default.
• W4310731609 cites W2007087804 @default.
• W4310731609 cites W2010789182 @default.
• W4310731609 cites W2017477261 @default.
• W4310731609 cites W2021823584 @default.
• W4310731609 cites W2029136954 @default.
• W4310731609 cites W2060021827 @default.
• W4310731609 cites W2067394538 @default.
• W4310731609 cites W2107318793 @default.
• W4310731609 cites W2107888488 @default.
• W4310731609 cites W2110780888 @default.
• W4310731609 cites W2116553639 @default.
• W4310731609 cites W2125728464 @default.
• W4310731609 cites W2143341110 @default.
• W4310731609 cites W2157559566 @default.
• W4310731609 cites W2278655101 @default.
• W4310731609 cites W2288377214 @default.
• W4310731609 cites W2330927521 @default.
• W4310731609 cites W2605432571 @default.
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• W4310731609 cites W2790606457 @default.
• W4310731609 cites W2796031506 @default.
• W4310731609 cites W2890647279 @default.
• W4310731609 cites W2892027944 @default.
• W4310731609 cites W2892794724 @default.
• W4310731609 cites W2947083497 @default.
• W4310731609 cites W2957183381 @default.
• W4310731609 cites W2965186303 @default.
• W4310731609 cites W2982498807 @default.
• W4310731609 cites W2995522849 @default.
• W4310731609 cites W3007131890 @default.
• W4310731609 cites W3012664562 @default.
• W4310731609 cites W3080151369 @default.
• W4310731609 cites W3091772497 @default.
• W4310731609 cites W3121132618 @default.
• W4310731609 cites W3137207553 @default.
• W4310731609 cites W3178136496 @default.
• W4310731609 cites W3179915633 @default.
• W4310731609 cites W4200292640 @default.
• W4310731609 cites W4200400903 @default.
• W4310731609 cites W4220933412 @default.
• W4310731609 cites W4280599430 @default.
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