A hybrid strategy combining solution NMR spectroscopy and isothermal titration calorimetry to characterize protein-nanodisc interaction
A hybrid strategy combining solution NMR spectroscopy and isothermal titration calorimetry to characterize protein-nanodisc interaction
复制标题
结合溶液核磁共振波谱和等温滴定量热法来表征蛋白质-纳米盘相互作用的混合策略
DOI：
10.1016/j.ab.2021.114521
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发表时间：
2022
影响因子：
2.9
通讯作者：
Kojima Chojiro
中科院分区：
文献类型：
--
作者：
Sugiki Toshihiko;Lee Young-Ho;Alsanousi Nesreen;Murata Kaito;Kawamura Izuru;Fujiwara Toshimichi;Hanada Kentaro;Kojima Chojiro
核磁共振是在原子分辨率上表征分子间相互作用的有力工具。然而，由于膜结合蛋白相互作用复杂的性质，使其相互作用机制难以阐明。在这里，我们证明了使用溶液核磁共振光谱和等温滴定量热法（ITC）的结构和热力学分析可以清楚地检测到神经酰胺转运蛋白（CERT）的pleckstrin同源（PH）结构域与嵌入在脂质纳米盘中的磷脂酰肌醇4-单磷酸（PI4P）之间的特异性相互作用，并区分与脂质纳米盘体积表面的特异性相互作用与非特异性相互作用。这种核磁共振- itc杂交策略提供了蛋白质-脂质膜相互作用的详细表征。
NMR is a powerful tool for characterizing intermolecular interactions at atomic resolution. However, the nature of the complex interactions of membrane-binding proteins makes it difficult to elucidate the interaction mechanisms. Here, we demonstrated that structural and thermodynamic analyses using solution NMR spectroscopy and isothermal titration calorimetry (ITC) can clearly detect a specific interaction between the pleckstrin homology (PH) domain of ceramide transport protein (CERT) and phosphatidylinositol 4-monophosphate (PI4P) embedded in the lipid nanodisc, and distinguish the specific interaction from nonspecific interactions with the bulk surface of the lipid nanodisc. This NMR-ITC hybrid strategy provides detailed characterization of protein-lipid membrane interactions.