The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster.
The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster.
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DOI：
10.1371/journal.pone.0003623
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发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Hutchings MI
中科院分区：
文献类型：
--
作者：
Tucker NP;Hicks MG;Clarke TA;Crack JC;Chandra G;Le Brun NE;Dixon R;Hutchings MI
调节蛋白NsrR是转录抑制因子Rrf 2家族的成员，专门用于感测多种致病性和非致病性细菌中的一氧化氮（NO）。有人提出NO通过与NsrR蛋白中预测的[Fe-S]簇相互作用直接调节NsrR活性，但没有实验证据支持这一假设。在这里，我们报告的纯化NsrR从专性需氧天蓝色链霉菌。我们证明，使用紫外可见，近紫外CD和EPR光谱，蛋白质含有NO敏感的[2Fe-2S]簇纯化时，从E。杆菌当NsrR暴露于NO时，该簇被亚硝基化，这导致DNA结合活性的丧失，如通过带移测定所检测的。去除[2Fe-2S]簇以产生apo-NsrR也导致DNA结合活性的丧失。这是第一次证明NsrR含有DNA结合活性所需的NO敏感的[2Fe-2S]簇。
The regulatory protein NsrR, a member of the Rrf2 family of transcription repressors, is specifically dedicated to sensing nitric oxide (NO) in a variety of pathogenic and non-pathogenic bacteria. It has been proposed that NO directly modulates NsrR activity by interacting with a predicted [Fe-S] cluster in the NsrR protein, but no experimental evidence has been published to support this hypothesis. Here we report the purification of NsrR from the obligate aerobe Streptomyces coelicolor. We demonstrate using UV-visible, near UV CD and EPR spectroscopy that the protein contains an NO-sensitive [2Fe-2S] cluster when purified from E. coli. Upon exposure of NsrR to NO, the cluster is nitrosylated, which results in the loss of DNA binding activity as detected by bandshift assays. Removal of the [2Fe-2S] cluster to generate apo-NsrR also resulted in loss of DNA binding activity. This is the first demonstration that NsrR contains an NO-sensitive [2Fe-2S] cluster that is required for DNA binding activity.
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影响因子：
2.9
作者：
Couture, MMJ;Colbert, CL;Eltis, LD
通讯作者：
Eltis, LD
DOI：
10.1073/pnas.1934677100
发表时间：
2003-11-25
影响因子：
11.1
作者：
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通讯作者：
Hopwood, DA
影响因子：
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作者：
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通讯作者：
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影响因子：
4.8
作者：
Kennedy, MC;Antholine, WE;Beinert, H
通讯作者：
Beinert, H
DOI：
10.1073/pnas.251550898
发表时间：
2001-12-18
影响因子：
11.1
作者：
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通讯作者：
Kiley, PJ