Probing domain interactions in soluble guanylate cyclase.
Probing domain interactions in soluble guanylate cyclase.
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DOI：
10.1021/bi200341b
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发表时间：
2011-05-24
期刊：
影响因子：
2.9
通讯作者：
Marletta, Michael A.
中科院分区：
文献类型：
--
作者：
Derbyshire, Emily R.;Winter, Michael B.;Ibrahim, Mohammed;Deng, Sarah;Spiro, Thomas G.;Marletta, Michael A.
真核一氧化氮（NO）信号转导涉及通过鸟苷酸环化酶（sGC）的可溶性同种型的活化来调节环GMP（cGMP）水平。sGC是一种异二聚体血红素蛋白，其含有血红素-一氧化氮和氧结合（H-NOX）结构域、Per/ARNT/Sim（PAS）结构域、卷曲螺旋（CC）结构域和催化结构域。为了评估这些结构域在调节NO敏感性sGC的血红素辅因子的配体结合特性中的作用，通过将大鼠β1 H-NOX结构域与来自腾冲嗜热厌氧杆菌、霍乱弧菌和秀丽隐杆线虫的含H-NOX结构域的蛋白质（分别为TtTar 4 H、VCA 0720和Gcy-33）的同源区域交换来构建嵌合体。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结构域影响细菌和蠕虫H-NOX结构域。cGMP生产在这些蛋白质的分析表明，含有细菌H-NOX的嵌合体表现出鸟苷酸环化酶活性，但这种活性不受气态配体结合血红素辅因子。含有非典型sGC Gcy-33 H-NOX结构域的大鼠蠕虫嵌合体被NO、CO和O2弱激活，表明非典型鸟苷酸环化酶和NO敏感性鸟苷酸环化酶具有共同的酶激活分子机制。为了探测其他sGC结构域对哺乳动物sGC血红素环境的影响，在β1 H-NOX构建体（残基1-194）、β1 H-NOX-PAS-CC构建体（残基1-385）和全长α1β1 sGC异二聚体（β1残基1-619）中产生血红素口袋突变体（Pro 118 Ala和Ile 145 Tyr）。这些蛋白质的光谱特性表明，域间通信调制血红素-NO复合物的配位状态和血红素氧化速率。总之，这些发现对含H-NOX蛋白的变构调节机制具有重要意义。
Eukaryotic nitric oxide (NO) signaling involves modulation in cyclic GMP (cGMP) levels through activation of the soluble isoform of guanylate cyclase (sGC). sGC is a heterodimeric hemoprotein that contains a Heme-Nitric oxide and OXygen binding (H-NOX) domain, a Per/ARNT/Sim (PAS) domain, a coiled-coil (CC) domain, and a catalytic domain. To evaluate the role of these domains in regulating the ligand binding properties of the heme cofactor of NO-sensitive sGC, chimeras were constructed by swapping the rat β1 H-NOX domain with the homologous region of H-NOX domain-containing proteins from Thermoanaerobacter tengcongensis, Vibrio cholerae, and Caenorhabditis elegans (TtTar4H, VCA0720, and Gcy-33, respectively). Characterization of ligand binding by electronic absorption and resonance Raman spectroscopy indicates that the other rat sGC domains influence the bacterial and worm H-NOX domains. Analysis of cGMP production in these proteins reveals that the chimeras containing bacterial H-NOXs exhibit guanylate cyclase activity, but this activity is not influenced by gaseous ligand binding to the heme cofactor. The rat-worm chimera containing the atypical sGC Gcy-33 H-NOX domain was weakly activated by NO, CO and O2, suggesting that atypical guanylate cyclases and NO-sensitive guanylate cyclases have a common molecular mechanism for enzyme activation. To probe the influence of the other sGC domains on the mammalian sGC heme environment, heme pocket mutants (Pro118Ala and Ile145Tyr) were generated in the β1 H-NOX construct (residues 1–194), the β1 H-NOX-PAS-CC construct (residues 1–385), and the full-length α1β1 sGC heterodimer (β1 residues 1–619). Spectroscopic characterization of these proteins shows that inter-domain communication modulates the coordination state of the heme-NO complex and the heme oxidation rate. Taken together, these findings have important implications for the allosteric mechanism of regulation within H-NOX containing proteins.
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影响因子：
2.9
作者：
Ibrahim, Mohammed;Derbyshire, Emily R.;Marletta, Michael A.;Spiro, Thomas G.
通讯作者：
Spiro, Thomas G.
影响因子：
11.4
作者：
Ma, Xiaolei;Sayed, Nazish;van den Akker, Focco
通讯作者：
van den Akker, Focco
影响因子：
2.9
作者：
Li, ZQ;Pal, B;Kitagawa, T
通讯作者：
Kitagawa, T
影响因子：
4.8
作者：
Derbyshire, Emily R.;Deng, Sarah;Marletta, Michael A.
通讯作者：
Marletta, Michael A.
DOI：
10.1016/b978-044452839-1.50005-x
发表时间：
2008-01-01
期刊：
SMALLEST BIOMOLECULES: DIATOMICS AND THEIR INTERACTIONS WITH HEME PROTEINS
影响因子：
--
作者：
Spiro, Thomas G.;Ibrahim, Mohammed;Wasbotten, Ingar H.
通讯作者：
Wasbotten, Ingar H.