Nitric Oxide and Heat Shock Protein 90 Activate Soluble Guanylate Cyclase by Driving Rapid Change in Its Subunit Interactions and Heme Content
Nitric Oxide and Heat Shock Protein 90 Activate Soluble Guanylate Cyclase by Driving Rapid Change in Its Subunit Interactions and Heme Content
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DOI：
10.1074/jbc.m114.559393
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发表时间：
2014-05-30
影响因子：
4.8
通讯作者：
Stuehr, Dennis J.
中科院分区：
文献类型：
--
作者：
Ghosh, Arnab;Stasch, Johannes-Peter;Stuehr, Dennis J.
背景：血红素插入可溶性鸟苷酸环化酶（sGC）使其能够结合一氧化氮（NO）进行细胞信号转导。结果：NO可诱导hsp 90依赖性血红素快速、可逆地插入sGC-1，并与sGC-1亚基结合。sGC激活剂BAY 60-2770也是如此。结论：NO动态影响活性sGC异源二聚体的成熟和稳定。重要性：这些数据揭示了调节细胞NO信号级联的新机制：分子伴侣热休克蛋白90（hsp 90）与细胞内的信号蛋白包括可溶性鸟苷酸环化酶（sGC）相关。hsp 90与无血红素（apo）sGC-1亚基结合，并有助于在sGC成熟为其NO响应活性形式期间驱动血红素插入。在此，我们发现，NO导致apo-sGC-1快速和短暂地从hsp 90解离并与细胞中的sGC-1缔合。这种NO反应（i）需要hsp 90是活性的，细胞血红素是可用的，并能够插入apo-sGC-1;（ii）与sGC-1血红素含量的增加有关;（iii）可以被血红素非依赖性sGC激活剂BAY 60-2770模拟;（iv）随后是sGC对NO的脱敏，sGC-1解离，并与hsp 90重新缔合。因此，NO促进快速、瞬时和hsp 90依赖性血红素插入细胞中的apo-sGC-1亚群中，这使其能够与sGC-1亚基联合收割机结合以形成成熟酶。驱动机制可能涉及sGC-1中血红素位点附近的构象变化，其可以被药理学sGC激活剂模拟。hsp 90、apo-sGC-1和sGC-1之间响应于NO的这种动态相互作用是前所未有的，并且代表了细胞可以调节sGC的血红素含量和活性以用于信号级联的新步骤。
Background: Heme insertion into souble guanylate cyclase (sGC) enables it to bind nitric oxide (NO) for cell signaling. Results: NO triggered a rapid, reversible, and hsp90-dependent heme insertion into sGC-1 and an association with sGC-1 subunit. sGC activator BAY 60-2770 did the same. Conclusion: NO dynamically impacts the maturation and stability of active sGC heterodimer. Significance: The data uncover new mechanisms that regulate cellular NO signaling cascades.The chaperone heat shock protein 90 (hsp90) associates with signaling proteins in cells including soluble guanylate cyclase (sGC). hsp90 associates with the heme-free (apo) sGC-1 subunit and helps to drive heme insertion during maturation of sGC to its NO-responsive active form. Here, we found that NO caused apo-sGC-1 to rapidly and transiently dissociate from hsp90 and associate with sGC-1 in cells. This NO response (i) required that hsp90 be active and that cellular heme be available and be capable of inserting into apo-sGC-1; (ii) was associated with an increase in sGC-1 heme content; (iii) could be mimicked by the heme-independent sGC activator BAY 60-2770; and (iv) was followed by desensitization of sGC toward NO, sGC-1 disassociation, and reassociation with hsp90. Thus, NO promoted a rapid, transient, and hsp90-dependent heme insertion into the apo-sGC-1 subpopulation in cells, which enabled it to combine with the sGC-1 subunit to form the mature enzyme. The driving mechanism likely involves conformational changes near the heme site in sGC-1 that can be mimicked by the pharmacologic sGC activator. Such dynamic interplay between hsp90, apo-sGC-1, and sGC-1 in response to NO is unprecedented and represent new steps by which cells can modulate the heme content and activity of sGC for signaling cascades.