Filling of a water-free void explains the allosteric regulation of the β1-adrenergic receptor by cholesterol
Filling of a water-free void explains the allosteric regulation of the β1-adrenergic receptor by cholesterol
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DOI：
10.1038/s41557-022-01009-9
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发表时间：
2022-08-11
期刊：
影响因子：
21.8
通讯作者：
Grzesiek, Stephan
中科院分区：
文献类型：
--
作者：
Abiko, Layara Akemi;Teixeira, Raphael Dias;Grzesiek, Stephan
最近的高压核磁共振结果表明，β（1）-肾上腺素能受体（β，AR）的预活性构象中存在体积为-100埃（3）的完全空的空腔，这些空腔在受体的活性构象中消失。在这里，我们使用氙衍生的β（1）AR晶体的X射线晶体学定位这些空腔。其中一个空腔与胆固醇结合口袋直接接触。溶液核磁共振表明，添加胆固醇类似物胆固醇半琥珀酸酯通过阻断保守的G蛋白偶联受体微型开关，阻碍了洗涤剂溶解的β（1）AR活性构象的形成，同时异丙肾上腺素和G蛋白模拟纳米抗体Nb 80对β（1）AR的亲和力降低。通过等温滴定量热法检测到AR。这种楔状作用解释了胆固醇作为β（1）AR负变构调节剂的功能。详细了解G蛋白偶联受体调节胆固醇通过填充干燥的空隙和容易侦察这样的空隙氙可能提供新的途径，为发展变构药物。
Recent high-pressure NMR results indicate that the preactive conformation of the beta(1)-adrenergic receptor (beta,AR) harbours completely empty cavities of -100 angstrom(3) volume, which disappear in the active conformation of the receptor. Here we have localized these cavities using X-ray crystallography of xenon-derivatized beta(1)AR crystals. One of the cavities is in direct contact with the cholesterol-binding pocket. Solution NMR shows that addition of the cholesterol analogue cholesteryl hemisuccinate impedes the formation of the active conformation of detergent-solubilized beta(1)AR by blocking conserved G protein-coupled receptor microswitches, concomitant with an affinity reduction of both isoprenaline and G protein-mimicking nanobody Nb80 for beta(1)AR detected by isothermal titration calorimetry. This wedge-like action explains the function of cholesterol as a negative allosteric modulator of beta(1)AR. A detailed understanding of G protein-coupled receptor regulation by cholesterol by filling of a dry void and the easy scouting for such voids by xenon may provide new routes for the development of allosteric drugs.