Structure/Activity Relationships of (M)ANT- and TNP-Nucleotides for Inhibition of Rat Soluble Guanylyl Cyclase α1β1
Structure/Activity Relationships of (M)ANT- and TNP-Nucleotides for Inhibition of Rat Soluble Guanylyl Cyclase α1β1
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DOI：
10.1124/mol.113.091017
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发表时间：
2014-04-01
影响因子：
3.6
通讯作者：
Seifert, Roland
中科院分区：
文献类型：
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作者：
Dove, Stefan;Danker, Kerstin Yvonne;Seifert, Roland
可溶性鸟苷酸环化酶(SGC)在心血管功能中起重要作用，并催化cGMP的形成。SGC被一氧化氮、变构刺激物和激活剂激活。然而，尽管与治疗相关，sGC的调节机制仍然不完全清楚。造成这种情况的一个主要原因是到目前为止还没有活性的sGC的晶体结构被解析。朝着这一目标迈出的重要一步是鉴定高亲和力的配体，这些配体能够稳定类似于活性的“完全封闭”状态的sGC构象。因此，我们检测了38个在核糖基端带有2，4，6，-三硝基苯基和(N-甲基)邻氨基苯甲酰基取代的嘌呤和嘧啶核苷酸对大鼠sGCα(1)β(1)的抑制作用，并将这些数据与结构相关的膜腺苷酸环化酶(MACs)1、2、5和纯化的Mac催化亚基Vc1：IIC2的数据进行了比较。TNP-GTP[2‘，3’-O-(2，4，6-三硝基苯基)-GTP]是最有效的sGCα(1)β(1)抑制剂(K-I，10.7 nM)，其次是2‘-mant-3’-dATP[2‘-O(N-甲基邻氨基苯甲酰基)-3’-脱氧-ATP](K-I，16.7 nM)。对来自催化结构域的非活性异二聚体晶体结构的sGCαCAT/sGCβCAT模型的对接研究表明，(M)ANT和TNP-核苷酸与sGCα(1)β(1)和Mac VC1：IIC2的作用相似。2‘-mant-3’-dATP和bi-(M)ANT-核苷酸在sGCα(1)β(1)的合理结合模式需要3‘-内切核糖构象(而在3’-mant-2‘-dATP中为3’-exo)。总体而言，sGCα(1)β(1)versusmacs 1、2、5的核苷酸抑制效力相关性较差。总体而言，我们确定了高效的sGCα(1)β(1)抑制剂，这些抑制剂可能对未来的结晶学和荧光光谱研究有用。此外，开发相对于sGC具有选择性的mac抑制剂是可能的。
Soluble guanylyl cyclase (sGC) plays an important role in cardiovascular function and catalyzes formation of cGMP. sGC is activated by nitric oxide and allosteric stimulators and activators. However, despite its therapeutic relevance, the regulatory mechanisms of sGC are still incompletely understood. A major reason for this situation is that no crystal structures of active sGC have been resolved so far. An important step toward this goal is the identification of high-affinity ligands that stabilize an sGC conformation resembling the active, "fully closed" state. Therefore, we examined inhibition of rat sGC alpha(1)beta(1) by 38 purine- and pyrimidine-nucleotides with 2,4,6,-trinitrophenyl and (N-methyl) anthraniloyl substitutions at the ribosyl moiety and compared the data with that for the structurally related membranous adenylyl cyclases (mACs) 1, 2, 5 and the purified mAC catalytic subunits VC1:IIC2. TNP-GTP [2',3'-O-(2,4,6-trinitrophenyl)-GTP] was the most potent sGC alpha(1)beta(1) inhibitor (K-i, 10.7 nM), followed by 2'-MANT-3'-dATP [2'-O(N-methylanthraniloyl)-3'-deoxy-ATP] (K-i, 16.7 nM). Docking studies on an sGC alpha cat/sGC beta cat model derived from the inactive heterodimeric crystal structure of the catalytic domains point to similar interactions of (M) ANT-and TNP-nucleotides with sGC alpha(1)beta(1) and mAC VC1:IIC2. Reasonable binding modes of 2'-MANT-3'-dATP and bis-(M)ANT-nucleotides at sGC alpha(1)beta(1) require a 3'-endo ribosyl conformation (versus 3'-exo in 3'-MANT-2'-dATP). Overall, inhibitory potencies of nucleotides at sGC alpha(1)beta(1) versusmACs 1, 2, 5 correlated poorly. Collectively, we identified highly potent sGC alpha(1)beta(1) inhibitors that may be useful for future crystallographic and fluorescence spectroscopy studies. Moreover, it may become possible to develop mAC inhibitors with selectivity relative to sGC.