Distinct molecular requirements for activation or stabilization of soluble guanylyl cyclase upon haem oxidation-induced degradation
Distinct molecular requirements for activation or stabilization of soluble guanylyl cyclase upon haem oxidation-induced degradation
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DOI：
10.1111/j.1476-5381.2009.00263.x
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发表时间：
2009-07-01
影响因子：
7.3
通讯作者：
Stasch, J. P.
中科院分区：
文献类型：
--
作者：
Hoffmann, L. S.;Schmidt, P. M.;Stasch, J. P.
背景和目的：在内皮功能障碍中，一氧化氮（NO）信号传导因可溶性鸟苷酸环化酶（sGC）血红素的氧化和随后的损失而受损。 sGC 激活剂 4-[((4-羧丁基){2-[(4-苯乙基苄基)氧基]苯乙基}氨基)甲基[苯甲酸]酸 (BAY 58-2667) 是一种血红素模拟物，当天然血红素（NO 结合位点）被去除时，能够以高亲和力与 sGC 结合，并且还可以保护 sGC 免受泛素触发的降解。在这里，我们研究这种保护是否是 BAY 58-2667 的独特特征，还是血红素位点配体的一般特征，例如不依赖血红素的 sGC 激活剂 5-氯-2-(5-氯-噻吩-2-磺酰氨基-N-(4-(吗啉-4-磺酰基)-苯基)-苯甲酰胺钠盐 (HMR 1766)、血红素模拟物Zn-原卟啉 IX (Zn-PPIX) 或血红素依赖性 sGC 刺激剂 5-环丙基-2-[1-(2-氟-苄基)-1H-吡唑并[3,4-b]吡啶-3-基]-嘧啶-4-基胺 (BAY 41-2272)。 实验方法：sGC 抑制剂1H-(1,2,4)-恶二唑并[4,3-a]喹喔啉-1-酮 (ODQ) 用于诱导氧化诱导的 sGC 降解。在中国仓鼠卵巢细胞系以及原代猪内皮细胞中测量了 sGC 的活性和蛋白水平，以阐明所观察到的影响的分子机制。相比之下，结构上不相关的 sGC 激活剂 HMR 1766 和 sGC 刺激剂 BAY 41-2272 没有保护作用，类似地，组成型无血红素的 sGC 突变体 beta(1)H105F 可以被 BAY 58-2667 稳定。 Zn-PPIX。结论：BAY 58-2667 不仅能够激活而且能够稳定氧化/无血红素 sGC，代表了双峰靶点相互作用的独特示例，并将该结构类别与非稳定 sGC 激活剂和 sGC 刺激剂（例如 HMR 1766 和 BAY 41-2272）区分开来。
Background and purpose:In endothelial dysfunction, signalling by nitric oxide (NO) is impaired because of the oxidation and subsequent loss of the soluble guanylyl cyclase (sGC) haem. The sGC activator 4-[((4-carboxybutyl){2-[(4-phenethylbenzyl)oxy]phenethyl}amino)methyl[benzoic]acid (BAY 58-2667) is a haem-mimetic able to bind with high affinity to sGC when the native haem (the NO binding site) is removed and it also protects sGC from ubiquitin-triggered degradation. Here we investigate whether this protection is a unique feature of BAY 58-2667 or a general characteristic of haem-site ligands such as the haem-independent sGC activator 5-chloro-2-(5-chloro-thiophene-2-sulphonylamino-N-(4-(morpholine-4-sulphonyl)-phenyl)-benzamide sodium salt (HMR 1766), the haem-mimetic Zn-protoporphyrin IX (Zn-PPIX) or the haem-dependent sGC stimulator 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-pyrimidin-4-ylamine (BAY 41-2272).Experimental approach:The sGC inhibitor 1H-(1,2,4)-oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) was used to induce oxidation-induced degradation of sGC. Activity and protein levels of sGC were measured in a Chinese hamster ovary cell line as well as in primary porcine endothelial cells. Cells expressing mutant sGC were used to elucidate the molecular mechanism underlying the effects observed.Key results:Oxidation-induced sGC degradation was prevented by BAY 58-2667 and Zn-PPIX in both cell types. In contrast, the structurally unrelated sGC activator, HMR 1766, and the sGC stimulator, BAY 41-2272, did not protect. Similarly, the constitutively haem-free sGC mutant beta(1)H105F was stabilized by BAY 58-2667 and Zn-PPIX.Conclusions:The ability of BAY 58-2667 not only to activate but also to stabilize oxidized/haem-free sGC represents a unique example of bimodal target interaction and distinguishes this structural class from non-stabilizing sGC activators and sGC stimulators such as HMR 1766 and BAY 41-2272, respectively.