Multi-region mapping of ligand binding and structural changes in the β-1 adrenergic receptor using carbene footprinting and mass spectrometry

IF 7.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Chemical Science Pub Date : 2025-10-01 DOI:10.1039/d5sc05107j

James Lloyd, Arppana Sara Varughese, Parth Kapoor, Katharina Duerr, Hsin-Yung Yen, Ali Jazayeri, Jonathan Hopper, Neil J Oldham

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引用次数: 0

Abstract

G protein coupled receptors (GPCRs) control many physiological processes and are major targets for therapeutic intervention. Transmembrane proteins, such as G protein–coupled receptors (GPCRs), are inherently flexible and dynamic and often challenging to study using conventional structural methods. Here, we report the use of carbene footprinting to investigate ligand binding and structural changes in the turkey β₁-adrenergic receptor (tβ₁AR), a GPCR and therapeutic target of beta-blocker drugs. The method revealed differences between binding of the agonist, isoprenaline, and the inverse agonist, carazalol, both in terms of their occupancy of the orthosteric ligand binding site and their effects on key regulatory structural features of tβ1AR including the ‘ionic lock’ between transmembrane (TM) helicies 3 and 6. Addition of nanobodies (Nbs) known to stabilise the activated complex (Nb80) and inactivated complex (Nb60) of tβ1AR induced further structural changes above those seen with the ligands alone.

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利用碳足迹和质谱技术对β-1肾上腺素能受体的配体结合和结构变化进行多区域定位

G蛋白偶联受体（gpcr）控制许多生理过程，是治疗干预的主要靶点。跨膜蛋白，如G蛋白偶联受体（gpcr），具有固有的灵活性和动态性，使用传统的结构方法研究往往具有挑战性。在这里，我们报告了使用碳足迹来研究火鸡β₁-肾上腺素能受体（tβ₁AR）的配体结合和结构变化，β₁AR是一种GPCR和β受体阻滞剂药物的治疗靶点。该方法揭示了激动剂异丙肾上腺素与逆激动剂卡拉唑尔的结合差异，这两种激动剂的结合方式都存在于正位配体结合位点上，以及它们对β 1ar关键调控结构特征的影响，包括跨膜（TM）螺旋3和6之间的“离子锁”。添加纳米体（Nbs）可以稳定t - β 1ar的活化复合物（Nb80）和失活复合物（Nb60），从而导致比单独使用配体所见的进一步的结构变化。

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来源期刊

Chemical Science CHEMISTRY, MULTIDISCIPLINARY-

CiteScore

14.40

自引率

4.80%

发文量

1352

审稿时长

2.1 months

期刊介绍： Chemical Science is a journal that encompasses various disciplines within the chemical sciences. Its scope includes publishing ground-breaking research with significant implications for its respective field, as well as appealing to a wider audience in related areas. To be considered for publication, articles must showcase innovative and original advances in their field of study and be presented in a manner that is understandable to scientists from diverse backgrounds. However, the journal generally does not publish highly specialized research.