Endogenous occurrence of protein S-guanylation in Escherichia coli: target identification and genetic regulation.
Endogenous occurrence of protein S-guanylation in Escherichia coli: target identification and genetic regulation.
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大肠杆菌中蛋白质 S-鸟苷酸化的内源发生：靶标识别和遗传调控。
DOI：
10.1016/j.bbrc.2016.07.110
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
T.
中科院分区：
文献类型：
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作者：
Tsutsuki;H.;Jung;M.;Zhang;T.;Ono;K.;Ida;T.;Kunieda;K.;Ihara;H.;Akaike;T. and Sawa;T.
8-硝基鸟苷3′，5 ′-环一磷酸（8-nitro-cGMP）是一种硝化cGMP衍生物，在一氧化氮（NO）和活性氧（ROS）的作用下形成。它可以通过cGMP加合（蛋白S-鸟苷酸化）引起蛋白巯基的翻译后修饰（PTM）。越来越多的证据表明，在哺乳动物中，氧化还原传感器蛋白的S-鸟苷酸化可能涉及对ROS相关氧化应激的适应性反应的调节。然而，细菌中S-鸟苷酸化的发生以及蛋白质靶点仍然未知。在这里，我们首次证明了大肠杆菌（E. coli）中蛋白质S-鸟苷化的内源性发生。用抗S-鸟苷酸抗体进行的Western印迹清楚地表明，在大肠杆菌中有多个蛋白质被S-鸟苷酸化。有趣的是，当细菌在静态培养条件下培养时，其中一些蛋白质比振荡培养条件下更强烈地S-鸟苷酸化。已知大肠杆菌缺乏鸟苷酸环化酶，鸟苷酸环化酶是哺乳动物中形成8-硝基-cGMP所必需的酶。我们发现大肠杆菌腺苷酸环化酶能催化前体8-硝基鸟苷5′-三磷酸生成8-硝基cGMP。更重要的是，E. colilacking腺苷酸环化酶显着减少S-鸟苷酸化蛋白的形成。我们的S-鸟苷酸化蛋白质组学成功鉴定了大肠杆菌中的S-鸟苷酸化蛋白质靶点，包括伴侣蛋白、核糖体蛋白以及与蛋白质合成、氧化还原调节和代谢相关的酶。了解蛋白质S-鸟苷酸化在细菌信号转导中的功能影响是开发潜在的化学治疗和新的诊断策略以控制病原性细菌感染的必要基础。
8-Nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP) is a nitrated cGMP derivative formed in response to nitric oxide (NO) and reactive oxygen species (ROS). It can cause a post-translational modification (PTM) of protein thiols through cGMP adduction (proteinS-guanylation). Accumulating evidence has suggested that, in mammals,S-guanylation of redox-sensor proteins may implicate in regulation of adaptive responses against ROS-associated oxidative stress. Occurrence as well as protein targets ofS-guanylation in bacteria remained unknown, however. Here we demonstrated, for the first time, the endogenous occurrence of proteinS-guanylation inEscherichia coli(E. coli). Western blotting using anti-S-guanylation antibody clearly showed that multiple proteins wereS-guanylated inE. coli. Interestingly, some of those proteins were more intenselyS-guanylated when bacteria were cultured under static culture condition than shaking culture condition. It has been known thatE. coliis deficient of guanylate cyclase, an enzyme indispensable for 8-nitro-cGMP formation in mammals. We found that adenylate cyclase fromE. colipotentially catalyzed 8-nitro-cGMP formation from its precursor 8-nitroguanosine 5′-triphosphate. More importantly,E. colilacking adenylate cyclase showed significantly reduced formation ofS-guanylated proteins. OurS-guanylation proteomics successfully identifiedS-guanylation protein targets inE. coli, including chaperons, ribosomal proteins, and enzymes which associate with protein synthesis, redox regulation and metabolism. Understanding of functional impacts for proteinS-guanylation in bacterial signal transduction is necessary basis for development of potential chemotherapy and new diagnostic strategy for control of pathogenic bacterial infections.