Hsp90 Regulates NADPH Oxidase Activity and Is Necessary for Superoxide but Not Hydrogen Peroxide Production
Hsp90 Regulates NADPH Oxidase Activity and Is Necessary for Superoxide but Not Hydrogen Peroxide Production
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DOI：
10.1089/ars.2010.3669
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发表时间：
2011-06-01
影响因子：
6.6
通讯作者：
Fulton, David J. R.
中科院分区：
文献类型：
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作者：
Chen, Feng;Pandey, Deepesh;Fulton, David J. R.
本研究的目的是确定热休克蛋白90 （Hsp90）是否调节NADPH氧化酶（Nox）的超氧化物和其他活性氧的产生。我们发现，Hsp90的药理学和遗传学抑制直接减少了nox5衍生的超氧化物，而不会二次改变信号事件。共免疫沉淀和生物发光共振能量转移研究表明，Nox5的c端与Hsp90结合。长期抑制Hsp90可降低Nox5的表达，进一步证明Nox5是Hsp90的客户蛋白。Hsp90抑制剂还能减少Nox1、Nox2（中性粒细胞）和Nox3的超氧化物。然而，仅释放过氧化氢的Nox4不受Hsp90抑制剂的影响。其他Nox酶的过氧化氢产量不受短期抑制Hsp90的影响，但长期抑制降低了所有活性氧的产量，同时酶的表达丧失。由n端Nox1或Nox3和c端Nox4组成的嵌合Nox酶的表达只产生对Hsp90抑制剂不敏感的过氧化氢。我们得出结论，Hsp90与Noxes1-3和5的c端结合，是酶稳定性和超氧化物产生所必需的。Hsp90不与Nox4的c端结合，也不需要过氧化氢的形成。Antioxid。氧化还原信号，14,2107-2119。
The goal of this study was to identify whether heat-shock protein 90 (Hsp90) regulates the production of superoxide and other reactive oxygen species from the NADPH oxidases (Nox). We found that pharmacological and genetic inhibition of Hsp90 directly reduced Nox5-derived superoxide without secondarily modifying signaling events. Coimmunoprecipitation and bioluminescence resonance energy transfer studies suggest that the C-terminus of Nox5 binds to Hsp90. Long-term Hsp90 inhibition reduced Nox5 expression and provides further evidence that Nox5 is an Hsp90 client protein. Inhibitors of Hsp90 also reduced superoxide from Nox1, Nox2 (neutrophils), and Nox3. However, Nox4, which emits only hydrogen peroxide, was unaffected by Hsp90 inhibitors. Hydrogen peroxide production from the other Nox enzymes was not affected by short-term inhibition of Hsp90, but long-term inhibition reduced production of all reactive oxygen species coincident with loss of enzyme expression. Expression of chimeric Nox enzymes consisting of N-terminal Nox1 or Nox3 and C-terminal Nox4 resulted in only hydrogen peroxide formation that was insensitive to Hsp90 inhibitors. We conclude that Hsp90 binds to the C-terminus of Noxes1-3 and 5 and is necessary for enzyme stability and superoxide production. Hsp90 does not bind to the C-terminus of Nox4 and is not required for hydrogen peroxide formation. Antioxid. Redox Signal. 14, 2107-2119.