Soluble guanylyl cyclase requires heat shock protein 90 for heme insertion during maturation of the NO-active enzyme
Soluble guanylyl cyclase requires heat shock protein 90 for heme insertion during maturation of the NO-active enzyme
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DOI：
10.1073/pnas.1205854109
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发表时间：
2012-08-07
影响因子：
11.1
通讯作者：
Stuehr, Dennis J.
中科院分区：
文献类型：
--
作者：
Ghosh, Arnab;Stuehr, Dennis J.
血红素插入是可溶性鸟苷酸环化酶（sGC）成熟过程中的关键，因为它使sGC能够识别NO并抑制其多种生物学效应。虽然sGC通常与细胞中的90 kDa热休克蛋白（hsp 90）相关，但其含义尚不清楚。目前的研究表明，热休克蛋白90是需要驱动血红素插入到sGC和完成其成熟。我们使用了哺乳动物细胞培养方法，并遵循血红素插入瞬时和内源性表达的无血红素sGC。我们使用药理学hsp 90抑制剂，ATP酶失活hsp 90突变体，血红素依赖性或血红素非依赖性sGC激活剂作为工具来破译hsp 90的作用。我们的研究结果表明，热休克蛋白90复合物apo-sGC，驱动血红素插入通过其固有的ATP酶活性，然后从成熟的，充满血红素的sGC解离。总之，这提高了我们对sGC成熟的理解，并揭示了控制细胞中sGC活性的独特方法，这对基于hsp 90的癌症治疗具有重要意义。
Heme insertion is key during maturation of soluble guanylyl cyclase (sGC) because it enables sGC to recognize NO and transduce its multiple biological effects. Although sGC is often associated with the 90-kDa heat shock protein (hsp90) in cells, the implications are unclear. The present study reveals that hsp90 is required to drive heme insertion into sGC and complete its maturation. We used a mammalian cell culture approach and followed heme insertion into transiently and endogenously expressed heme-free sGC. We used pharmacological hsp90 inhibitors, an ATP-ase inactive hsp90 mutant, and heme-dependent or heme-independent sGC activators as tools to decipher the role of hsp90. Our findings suggest that hsp90 complexes with apo-sGC, drives heme insertion through its inherent ATPase activity, and then dissociates from the mature, heme-replete sGC. Together, this improves our understanding of sGC maturation and reveals a unique means to control sGC activity in cells, and it has important implications for hsp90 inhibitor-based cancer therapy.