Mapping the Endothelial Cell S-Sulfhydrome Highlights the Crucial Role of Integrin Sulfhydration in Vascular Function
Mapping the Endothelial Cell S-Sulfhydrome Highlights the Crucial Role of Integrin Sulfhydration in Vascular Function
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DOI：
10.1161/circulationaha.120.051877
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发表时间：
2021-03-02
期刊：
影响因子：
37.8
通讯作者：
Fleming, Ingrid
中科院分区：
文献类型：
--
作者：
Bibli, Sofia-Iris;Hu, Jiong;Fleming, Ingrid
背景资料：在血管内皮细胞中，通过胱硫醚γ裂解酶（CSE）的半胱氨酸代谢产生硫化氢相关的硫烷硫化合物（H2Sn），其通过靶蛋白的半胱氨酸S-硫水合作用发挥其生物学作用。方法：采用液相色谱-串联质谱法（LC-MS-MS）对人内皮细胞蛋白质中的S-巯基半胱氨酸和β 3整合素蛋白质内二硫键重排进行鉴定。功能研究包括内皮细胞粘附，剪切应力诱导的细胞排列，血压测量，以及在内皮细胞特异性CSE基因敲除小鼠和一小部分内皮功能障碍患者中流动诱导的血管舒张。（1）从无斑块肠系膜动脉分离的天然人内皮细胞（CSE活性高）和含斑块的颈动脉（2）培养的人内皮细胞保持在静态条件下或暴露于流体剪切应力以降低CSE表达;和（3）培养的内皮细胞暴露于剪切应力以降低CSE表达，并用溶剂或缓释H2Sn供体SG 1002处理。内皮细胞“S-巯基”由1591个蛋白质中的3446个单独的半胱氨酸残基组成。改变最多的蛋白质家族是整合素，详细关注β 3整合素，我们发现S-巯基化影响蛋白质内二硫键的形成，并且是维持β腿的扩展开放构象所必需的。β 3整联蛋白S-硫化是体外内皮细胞机械转导以及鼠肠系膜动脉中流动诱导的扩张所需的。在培养的细胞中，S-巯基化的丧失损害了β 3整合素和G α 13（鸟嘌呤核苷酸结合蛋白亚基α 13）之间的相互作用，导致RhoA（ras同源家族成员A）的组成性激活，并损害了血流诱导的内皮细胞重新排列。在人类动脉粥样硬化，内皮功能与低H2Sn的产生，受损的流量诱导的扩张，并未能检测β 3整合素S-sulfhydration，所有这些都获救后，管理的H2Sn supplement.Conclusions：血管疾病与显着变化的S-sulfhydration的内皮细胞蛋白参与介导的流量。短期补充H2Sn改善了人类的血管反应性，突出了干扰这一途径治疗血管疾病的潜力。
Background: In vascular endothelial cells, cysteine metabolism by the cystathionine gamma lyase (CSE), generates hydrogen sulfide-related sulfane sulfur compounds (H2Sn), that exert their biological actions via cysteine S-sulfhydration of target proteins. This study set out to map the "S-sulfhydrome" (ie, the spectrum of proteins targeted by H2Sn) in human endothelial cells.Methods: Liquid chromatography with tandem mass spectrometry was used to identify S-sulfhydrated cysteines in endothelial cell proteins and beta 3 integrin intraprotein disulfide bond rearrangement. Functional studies included endothelial cell adhesion, shear stress-induced cell alignment, blood pressure measurements, and flow-induced vasodilatation in endothelial cell-specific CSE knockout mice and in a small collective of patients with endothelial dysfunction.Results: Three paired sample sets were compared: (1) native human endothelial cells isolated from plaque-free mesenteric arteries (CSE activity high) and plaque-containing carotid arteries (CSE activity low); (2) cultured human endothelial cells kept under static conditions or exposed to fluid shear stress to decrease CSE expression; and (3) cultured endothelial cells exposed to shear stress to decrease CSE expression and treated with solvent or the slow-releasing H2Sn donor, SG1002. The endothelial cell "S-sulfhydrome" consisted of 3446 individual cysteine residues in 1591 proteins. The most altered family of proteins were the integrins and focusing on beta 3 integrin in detail we found that S-sulfhydration affected intraprotein disulfide bond formation and was required for the maintenance of an extended-open conformation of the beta leg. beta 3 integrin S-sulfhydration was required for endothelial cell mechanotransduction in vitro as well as flow-induced dilatation in murine mesenteric arteries. In cultured cells, the loss of S-sulfhydration impaired interactions between beta 3 integrin and G alpha 13 (guanine nucleotide-binding protein subunit alpha 13), resulting in the constitutive activation of RhoA (ras homolog family member A) and impaired flow-induced endothelial cell realignment. In humans with atherosclerosis, endothelial function correlated with low H2Sn generation, impaired flow-induced dilatation, and failure to detect beta 3 integrin S-sulfhydration, all of which were rescued after the administration of an H2Sn supplement.Conclusions: Vascular disease is associated with marked changes in the S-sulfhydration of endothelial cell proteins involved in mediating responses to flow. Short-term H2Sn supplementation improved vascular reactivity in humans highlighting the potential of interfering with this pathway to treat vascular disease.