Discovering protein-protein interaction stabilisers by native mass spectrometry.
Discovering protein-protein interaction stabilisers by native mass spectrometry.
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DOI：
10.1039/d1sc01450a
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发表时间：
2021-08-18
期刊：
影响因子：
8.4
通讯作者：
Leney AC
中科院分区：
文献类型：
--
作者：
Bellamy-Carter J;Mohata M;Falcicchio M;Basran J;Higuchi Y;Doveston RG;Leney AC
蛋白质-蛋白质相互作用（PPI）是关键的治疗靶点。临床上大多数 PPI 靶向药物都会抑制这些重要的相互作用；然而，在 PPI 在疾病状态下受到干扰的情况下，稳定 PPI 是一种有吸引力的替代方案。由于缺乏可用于监测 PPI 稳定性的工具，新型 PPI 稳定剂的发现受到阻碍。此外，为了检测 PPI 稳定性，需要同时监测结合的化学计量及其对结合平衡的变化。在这里，我们展示了原生质谱 (MS) 在快速寻找 PPI 稳定剂方面的强大功能。为了证明其能力，我们重点研究了真核调节蛋白 14-3-3σ 及其结合伙伴雌激素受体 ERα、肿瘤抑制因子 p53 和激酶 LRRK2 之间的三个 PPI，它们在添加小分子 Fusicoccin A 后的相互作用得到了差异稳定。在单次测量中，14-3-3 与其每个结合配偶体之间的化学计量和结合平衡是明显的。添加梭菌素 A 稳定剂后，与 14-3-3:p53 和 14-3-3:LRRK2 复合物相比，14-3-3:ERα 复合物的结合平衡发生显着变化。我们的结果强调了天然 MS 不仅能够区分稳定剂调节 PPI 的能力，而且还能为三元复合物形成的动力学提供重要的见解。最后，我们展示了如何使用天然 MS 作为寻找 PPI 稳定剂的筛选工具，强调其作为寻找新型治疗性 PPI 稳定剂的初级筛选技术的潜在作用。稳定蛋白质-蛋白质相互作用具有挑战性，但在治疗上很重要。天然质谱可用于监测结合平衡，从而识别和测量新型蛋白质-蛋白质相互作用稳定剂。
Protein–protein interactions (PPIs) are key therapeutic targets. Most PPI-targeting drugs in the clinic inhibit these important interactions; however, stabilising PPIs is an attractive alternative in cases where a PPI is disrupted in a disease state. The discovery of novel PPI stabilisers has been hindered due to the lack of tools available to monitor PPI stabilisation. Moreover, for PPI stabilisation to be detected, both the stoichiometry of binding and the shift this has on the binding equilibria need to be monitored simultaneously. Here, we show the power of native mass spectrometry (MS) in the rapid search for PPI stabilisers. To demonstrate its capability, we focussed on three PPIs between the eukaryotic regulatory protein 14-3-3σ and its binding partners estrogen receptor ERα, the tumour suppressor p53, and the kinase LRRK2, whose interactions upon the addition of a small molecule, fusicoccin A, are differentially stabilised. Within a single measurement the stoichiometry and binding equilibria between 14-3-3 and each of its binding partners was evident. Upon addition of the fusicoccin A stabiliser, a dramatic shift in binding equilibria was observed with the 14-3-3:ERα complex compared with the 14-3-3:p53 and 14-3-3:LRRK2 complexes. Our results highlight how native MS can not only distinguish the ability of stabilisers to modulate PPIs, but also give important insights into the dynamics of ternary complex formation. Finally, we show how native MS can be used as a screening tool to search for PPI stabilisers, highlighting its potential role as a primary screening technology in the hunt for novel therapeutic PPI stabilisers. Stabilising protein–protein interactions is challenging, yet therapeutically important. Native mass spectrometry can be used to monitor binding equilibria, allowing identification and measurement of novel protein–protein interaction stabilisers.
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影响因子：
4.3
作者：
Bosica, Francesco;Andrei, Sebastian A.;O'Mahony, Gavin
通讯作者：
O'Mahony, Gavin
影响因子：
14.8
作者：
Hernandez, Helena;Robinson, Carol V.
通讯作者：
Robinson, Carol V.
DOI：
10.1042/bj20091834
发表时间：
2010-03-15
期刊：
The Biochemical journal
影响因子：
--
作者：
Johnson C;Crowther S;Stafford MJ;Campbell DG;Toth R;MacKintosh C
通讯作者：
MacKintosh C
影响因子：
7
作者：
Falcicchio M;Ward JA;Macip S;Doveston RG
通讯作者：
Doveston RG
影响因子：
3.5
作者：
Doveston, Richard G.;Kuusk, Ave;Ottmann, Christian
通讯作者：
Ottmann, Christian