S-nitrosylation of Hsp90 promotes the inhibition of its ATPase and endothelial nitric oxide synthase regulatory activities
S-nitrosylation of Hsp90 promotes the inhibition of its ATPase and endothelial nitric oxide synthase regulatory activities
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DOI：
10.1073/pnas.0407294102
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发表时间：
2005-06-14
影响因子：
11.1
通讯作者：
Lamas, S
中科院分区：
文献类型：
--
作者：
Martínez-Ruiz, A;Villanueva, L;Lamas, S
一氧化氮参与不同系统的多种信号通路，尤其是内皮细胞。它的一些作用可以通过蛋白质的共价修饰来发挥，在这些修饰中，越来越多的人关注s -亚硝基化作为一种信号机制。在这项工作中，我们通过多种方法（臭氧化学发光，生物素开关和质谱）证明分子伴侣Hsp90是s -亚硝基化的靶标，并在c端区域鉴定出与内皮型一氧化氮合酶（eNOS）相互作用的易感半胱氨酸残基。我们还发现，内皮细胞在接受s -亚硝基- l -半胱氨酸处理和暴露于eNOS激活剂时也会发生这种修饰。Hsp90 atp酶活性及其对eNOS活性的积极作用均被s -亚硝基化抑制。总之，这些数据表明，s -亚硝基化可能在功能上调节Hsp90的一般活性，并为限制eNOS激活提供反馈机制。
Nitric oxide is implicated in a variety of signaling pathways in different systems, notably in endothelial cells. Some of its effects can be exerted through covalent modifications of proteins and, among these modifications, increasing attention is being paid to S-nitrosylation as a signaling mechanism. In this work, we show by a variety of methods (ozone chemiluminescence, biotin switch, and mass spectrometry) that the molecular chaperone Hsp90 is a target of S-nitrosylation and identify a susceptible cysteine residue in the region of the C-terminal domain that interacts with endothelial nitric oxide synthase (eNOS). We also show that the modification occurs in endothelial cells when they are treated with S-nitroso-L-Cysteine and when they are exposed to eNOS activators. Hsp90 ATPase activity and its positive effect on eNOS activity are both inhibited by S-nitrosylation. Together, these data suggest that S-nitrosylation may functionally regulate the general activities of Hsp90 and provide a feedback mechanism for limiting eNOS activation.