...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Probing Domain Interactions in Soluble Guanylate Cyclase
【24h】

Probing Domain Interactions in Soluble Guanylate Cyclase

机译：探索可溶性鸟苷酸环化酶中的域相互作用

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Eukaryotic nitric oxide (NO) signaling involves modulation of cyclic GMP (cGMP) levelsnthrough activation of the soluble isoform of guanylate cyclase (sGC). sGC is a heterodimericnhemoprotein that contains a Hemeu0002Nitric oxide and OXygen binding (H-NOX) domain, a PerARNT/Sim (PAS) domain, a coiled-coil (CC) domain, and a catalytic domain. To evaluate the role ofnthese domains in regulating the ligand binding properties of the heme cofactor of NO-sensitive sGC, wenconstructed chimeras by swapping the rat β1 H-NOX domain with the homologous region of H-NOXndomain-containing proteins from Thermoanaerobacter tengcongensis, Vibrio cholerae, and Caenorhabditisnelegans (TtTar4H, VCA0720, and Gcy-33, respectively). Characterization of ligand binding by electronicnabsorption and resonance Raman spectroscopy indicates that the other rat sGC domains influence thenbacterial and worm H-NOX domains. Analysis of cGMP production in these proteins reveals that thenchimeras containing bacterial H-NOX domains exhibit guanylate cyclase activity, but this activity is notninfluenced by gaseous ligand binding to the heme cofactor. The ratu0002worm chimera containing thenatypical sGC Gcy-33 H-NOX domain was weakly activated by NO, CO, and O2, suggesting that atypicalnguanylate cyclases and NO-sensitive guanylate cyclases have a common molecular mechanism for enzyme activation. To probe theninfluence of the other sGC domains on the mammalian sGC heme environment, we generated heme pocket mutants (Pro118Alanand Ile145Tyr) in the β1 H-NOX construct (residues 1u0002194), the β1 H-NOX-PAS-CC construct (residues 1u0002385), and the full-nlength R1β1 sGC heterodimer (β1 residues 1u0002619). Spectroscopic characterization of these proteins shows that interdomainncommunication modulates the coordination state of the hemeu0002NO complex and the heme oxidation rate. Taken together, thesenfindings have important implications for the allosteric mechanism of regulation within H-NOX domain-containing proteins.
机译：真核一氧化氮（NO）信号传导涉及通过激活鸟苷酸环化酶（sGC）的可溶性同工型来调节环状GMP（cGMP）水平。 sGC是一种异质二聚体血红蛋白，包含Hemeu0002一氧化氮和氧结合（H-NOX）域，Per / nARNT / Sim（PAS）域，卷曲螺旋（CC）域和催化域。为了评估这些域在调节NO敏感sGC血红素辅因子的配体结合特性中的作用，通过将大鼠β1H-NOX域与来自滕氏嗜热厌氧杆菌，霍乱弧菌的含H-NOXn域的蛋白质的同源区域交换，构建了嵌合体。 ，和Caenorhabditisnelegans（分别为TtTar4H，VCA0720和Gcy-33）。通过电子吸收和共振拉曼光谱表征配体结合，表明其他大鼠sGC域影响细菌和蠕虫H-NOX域。对这些蛋白质中cGMP产生的分析表明，含有细菌H-NOX结构域的羊膜显示了鸟苷酸环化酶活性，但是这种气态配体与血红素辅因子的结合并没有影响。含有典型sGC Gcy-33 H-NOX结构域的ratu0002worm嵌合体被NO，CO和O2弱激活，表明非典型鸟苷酸环化酶和NO敏感鸟苷酸环化酶具有共同的酶激活分子机制。为了探查其他sGC域对哺乳动物sGC血红素环境的影响，我们在β1H-NOX构建体（残基1u0002194），β1H-NOX-PAS-CC构建体（残基1u0002385）中生成了血红素口袋突变体（Pro118Alanand Ile145Tyr）。 ，以及全长R1β1sGC异二聚体（β1残基1u0002619）。这些蛋白质的光谱表征表明，域间通信调节了hemeu0002NO复合物的配位状态和血红素的氧化速率。两者合计，这些发现对含H-NOX域的蛋白质的变构调节机制具有重要意义。

著录项

  • 来源
    《Biochemistry》 |2011年第20期|p.4281-4290|共10页
  • 作者

    Emily R. Derbyshire Michael B. Winter Mohammed Ibrahim Sarah Deng Thomas G. Spiro and Michael A. Marletta;

  • 作者单位

    †Department of Molecular and Cell Biology,‡Department of Chemistry, and §Department of Plant and Microbial Biology,University of California, Berkeley, California 94720, United States) Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, United States^California Institute for Quantitative Biosciences and @Division of Physical Biosciences, University of California, Berkeley,California 94720-3220, United States;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号