GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain.
GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain.
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DOI：
10.1073/pnas.2108079118
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发表时间：
2021-08-03
影响因子：
11.1
通讯作者：
Wellendorph P
中科院分区：
文献类型：
--
作者：
Leurs U;Klein AB;McSpadden ED;Griem-Krey N;Solbak SMØ;Houlton J;Villumsen IS;Vogensen SB;Hamborg L;Gauger SJ;Palmelund LB;Larsen ASG;Shehata MA;Kelstrup CD;Olsen JV;Bach A;Burnie RO;Kerr DS;Gowing EK;Teurlings SMW;Chi CC;Gee CL;Frølund B;Kornum BR;van Woerden GM;Clausen RP;Kuriyan J;Clarkson AN;Wellendorph P
GHB是GABA的天然脑代谢物，先前报道具有神经保护作用。然而，GHB的高亲和力结合位点在近40年来一直难以捉摸。我们在这里揭示了CaMKIIα，一种非常重要的神经元激酶，作为长期寻求的GHB高亲和力靶点。通过CaMKIIα中心枢纽结构域内的特异性相互作用，GHB类似物起到稳定枢纽寡聚体复合物的作用。这种相互作用可能解释了GHB类似物在暴露于化学损伤的培养神经元和暴露于缺血的小鼠中的显著神经保护作用。GHB类似物的缺血后治疗作用强调了这些化合物作为选择性和高亲和力的潜在候选药物以及CaMKIIα作为中风治疗的相关药理学靶点。Ca 2 +/钙调蛋白依赖性蛋白激酶II α亚基（CaMKIIα）是一种关键的神经元信号蛋白，也是一种新兴的药物靶点。中枢结构域通过将全酶复合物组织成功能性寡聚体来调节CaMK II α的活性，但中枢结构域的药理学调节从未得到证实。在这里，使用光亲和标记和化学蛋白质组学的组合，我们表明与天然物质γ-羟基丁酸酯（GHB）相关的化合物选择性地结合CaMKIIα。通过配体结合的CaMKIIα枢纽的2.2-X射线晶体结构，我们揭示了枢纽深处结合位点的分子细节。此外，我们表明，GHB和相关类似物结合到这个网站促进集线器热稳定性的浓度依赖性增加，据信改变全酶功能。在病理性CaMKIIα激活状态下，中枢配体选择性地提供显著和持续的神经保护，这是时间和剂量依赖性的。这在暴露于兴奋性毒性的神经元和使用选择性GHB类似物HOCPCA（3-羟基环己-1-烯羧酸）的脑缺血小鼠模型中得到证实。总之，我们的研究结果表明，迄今为止未知的神经保护机制，通过高度特异性和不可预见的相互作用之间的CaMKIIα枢纽结构域和小分子脑渗透GHB类似物。这确立了GHB类似物作为研究CaMKII神经药理学的有力工具，特别是作为脑缺血的潜在治疗化合物。
GHB is a natural brain metabolite of GABA, previously reported to be neuroprotective. However, the high-affinity binding site for GHB has remained elusive for almost 40 y. We here unveil CaMKIIα, a highly important neuronal kinase, as the long-sought-after GHB high-affinity target. Via a specific interaction within the central hub domain of CaMKIIα, GHB analogs act to stabilize the hub oligomer complex. This interaction potentially explains pronounced neuroprotective effects of GHB analogs in cultured neurons exposed to a chemical insult and in mice exposed to ischemia. The postischemic treatment effects of GHB analogs underline these compounds as selective and high-affinity potential drug candidates and CaMKIIα as a relevant pharmacological target for stroke therapy. Ca2+/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) is a key neuronal signaling protein and an emerging drug target. The central hub domain regulates the activity of CaMKIIα by organizing the holoenzyme complex into functional oligomers, yet pharmacological modulation of the hub domain has never been demonstrated. Here, using a combination of photoaffinity labeling and chemical proteomics, we show that compounds related to the natural substance γ-hydroxybutyrate (GHB) bind selectively to CaMKIIα. By means of a 2.2-Å x-ray crystal structure of ligand-bound CaMKIIα hub, we reveal the molecular details of the binding site deep within the hub. Furthermore, we show that binding of GHB and related analogs to this site promotes concentration-dependent increases in hub thermal stability believed to alter holoenzyme functionality. Selectively under states of pathological CaMKIIα activation, hub ligands provide a significant and sustained neuroprotection, which is both time and dose dependent. This is demonstrated in neurons exposed to excitotoxicity and in a mouse model of cerebral ischemia with the selective GHB analog, HOCPCA (3-hydroxycyclopent-1-enecarboxylic acid). Together, our results indicate a hitherto unknown mechanism for neuroprotection by a highly specific and unforeseen interaction between the CaMKIIα hub domain and small molecule brain-penetrant GHB analogs. This establishes GHB analogs as powerful tools for investigating CaMKII neuropharmacology in general and as potential therapeutic compounds for cerebral ischemia in particular.
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影响因子：
8.2
作者：
Coultrap, Steven J.;Vest, Rebekah S.;Ashpole, Nicole M.;Hudmon, Andy;Bayer, K. Ulrich
通讯作者：
Bayer, K. Ulrich
影响因子：
7.7
作者：
Karandur D;Bhattacharyya M;Xia Z;Lee YK;Muratcioglu S;McAffee D;McSpadden ED;Qiu B;Groves JT;Williams ER;Kuriyan J
通讯作者：
Kuriyan J
影响因子：
56.9
作者：
De Koninck, P;Schulman, H
通讯作者：
Schulman, H
影响因子：
7.2
作者：
Bhattacharyya, Moitrayee;Karandur, Deepti;Kuriyan, John
通讯作者：
Kuriyan, John
影响因子：
7.7
作者：
Chia PH;Zhong FL;Niwa S;Bonnard C;Utami KH;Zeng R;Lee H;Eskin A;Nelson SF;Xie WH;Al-Tawalbeh S;El-Khateeb M;Shboul M;Pouladi MA;Al-Raqad M;Reversade B
通讯作者：
Reversade B