Luminescence resonance energy transfer spectroscopy of ATP-binding cassette proteins.
Luminescence resonance energy transfer spectroscopy of ATP-binding cassette proteins.
复制标题
ATP 结合盒蛋白的发光共振能量转移光谱。
DOI：
10.1016/j.bbamem.2017.08.005
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Altenberg,GuillermoA
中科院分区：
文献类型：
--
作者：
Zoghbi,MariaE;Altenberg,GuillermoA
ATP结合盒（ABC）超家族包括调节和转运蛋白。大多数人类ABC输出者利用ATP水解产生的能量将底物泵出细胞。虽然在理解ABC输出者的分子机制方面取得了重大进展，但仍有许多问题尚未解决。在过去的几年中，发光共振能量转移已被用于检测构象变化，在真实的时间，原子分辨率，在孤立的ABC核苷酸结合域（NBD）和全长ABC出口商。NBD是特别令人感兴趣的，因为它们提供了用于基板传输的动力行程。发光共振能量转移（LRET）是一种光谱技术，可以提供蛋白质在生理条件下构象变化的原子分辨率的动态信息。使用LRET，已经表明NBD二聚化，ABC蛋白催化循环中的关键步骤，需要ATP结合到两个核苷酸结合位点。然而，仅在其中一个位点处的水解可驱动NBD二聚体的解离。还发现，在脂质双层膜中重构并在37 °C下研究的细菌ABC输出者MsbA的NBD从未像晶体结构所表明的那样分离。这一观察强调了在生理条件下进行ABC出口商的结构/功能研究的重要性。本文是由Ute Hellmich，Rupak Doshi和Benjamin McIlmet编辑的题为：超越膜蛋白质的结构-功能视野的特刊的一部分。
The ATP-binding cassette (ABC) superfamily includes regulatory and transport proteins. Most human ABC exporters pump substrates out of cells using energy from ATP hydrolysis. Although major advances have been made toward understanding the molecular mechanism of ABC exporters, there are still many issues unresolved. During the last few years, luminescence resonance energy transfer has been used to detect conformational changes in real time, with atomic resolution, in isolated ABC nucleotide binding domains (NBDs) and full-length ABC exporters. NBDs are particularly interesting because they provide the power stroke for substrate transport. Luminescence resonance energy transfer (LRET) is a spectroscopic technique that can provide dynamic information with atomic-resolution of protein conformational changes under physiological conditions. Using LRET, it has been shown that NBD dimerization, a critical step in ABC proteins catalytic cycle, requires binding of ATP to two nucleotide binding sites. However, hydrolysis at just one of the sites can drive dissociation of the NBD dimer. It was also found that the NBDs of the bacterial ABC exporter MsbA reconstituted in a lipid bilayer membrane and studied at 37 °C never separate as much as suggested by crystal structures. This observation stresses the importance of performing structural/functional studies of ABC exporters under physiologic conditions. This article is part of a Special Issue entitled: Beyond the Structure-Function Horizon of Membrane Proteins edited by Ute Hellmich, Rupak Doshi and Benjamin McIlwain.