Hsp90 S-nitrosylation at Cys521, as a conformational switch, modulates cycling of Hsp90-AHA1-CDC37 chaperone machine to aggravate atherosclerosis

• Published in: Redox biology 2022-06, Vol.52, p.102290-102290, Article 102290
• Main Authors: Zhao, Shuang, Tang, Xin, Miao, Zian, Chen, Yurong, Cao, Jiawei, Song, Tianyu, You, Daiting, Zhong, Yanqing, Lin, Zhe, Wang, Dan, Shi, Zhiguang, Tang, Xinlong, Wang, Dongjin, Chen, Shaoliang, Wang, Liansheng, Gu, Aihua, Chen, Feng, Xie, Liping, Huang, Zhengrong, Wang, Hong, Ji, Yong
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphatases; Animals; Atherosclerosis; Atherosclerosis - genetics; Atherosclerosis - metabolism; Chromatography, Liquid; Cysteine - metabolism; Endothelial Cells - metabolism; Endothelial dysfunction; Heat shock protein 90; HSP90 Heat-Shock Proteins - genetics; HSP90 Heat-Shock Proteins - metabolism; Mice; Molecular Chaperones - metabolism; Nitric Oxide Synthase Type III - genetics; Nitric Oxide Synthase Type III - metabolism; Oxidative stress; Research Paper; S-nitrosylation; Tandem Mass Spectrometry
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2022.102290

Endothelial dysfunction is the initial process of atherosclerosis. Heat shock protein 90 (Hsp90), as a molecular chaperone, plays a crucial role in various cardiovascular diseases. Hsp90 function is regulated by S-nitrosylation (SNO). However, the precise role of SNO-Hsp90 in endothelial dysfunction during atherosclerosis remains unclear. We here identified Hsp90 as a highly S-nitrosylated target in endothelial cells (ECs) by biotin switch assay combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The elevation of SNO-Hsp90 was observed in atherosclerotic human and rodent aortas as well as in oxidized LDL (oxLDL)-treated ECs. Inhibition of inducible nitric oxide synthase (iNOS) or transfection with Hsp90 cysteine 521 (Cys521) mutation plasmid decreased the level of SNO-Hsp90 in oxLDL-cultured ECs. Coimmunoprecipitation and proximity ligation assay demonstrated that SNO-Hsp90 at Cys521 suppressed the interaction between Hsp90 and activator of Hsp90 ATPase activity 1 (AHA1), but promoted the association of Hsp90 and cell division cycle 37 (CDC37). Hsp90 Cys521 mutation increased endothelial nitric oxide synthase (eNOS) activity and inhibited nuclear factor kappa-B (NF-κB) signaling, thereby increasing nitric oxide (NO) bioavailability and alleviating endothelial adhesion, inflammation and oxidative stress in oxLDL-treated ECs. Also, administration of endothelial-specific adeno-associated viruses of Cys521-mutated Hsp90 significantly mitigated vascular oxidative stress, macrophage infiltration and atherosclerosis lesion areas in high fat diet-fed ApoE-/- mice. In conclusion, SNO-Hsp90 at Cys521, that serves as a conformational switch, disrupts Hsp90/AHA1 interaction but promotes recruitment of CDC37 to exacerbate atherosclerosis. [Display omitted] •Hsp90 S-nitrosylation at Cys521 acts as a conformational switch to modulate Hsp90/AHA1 and Hsp90/CDC37 interaction.•SNO-Hsp90 induces endothelial adhesion, inflammation and oxidative stress.•SNO-Hsp90 mediates endothelial dysfunction to exacerbate atherosclerosis.