Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B.
Room-temperature serial synchrotron crystallography of the human phosphatase PTP1B.
复制标题
DOI：
10.1107/s2053230x22011645
复制
发表时间：
2023-01-01
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
第一个系列的同步晶体学结构的载脂蛋白PTP1B的报告和构象异质性在几个变构网站相比，在以前的结构。室温X射线晶体学为蛋白质构象异质性提供了独特的见解，但获得足够大的蛋白质晶体是一个常见的障碍。连续同步加速器晶体学（SSX）通过允许使用许多中到小尺寸的晶体来帮助解决这个障碍。在这里，最近推出的连续样品支持芯片系统已被用来获得第一个SSX结构的人磷酸酶，特别是蛋白酪氨酸磷酸酶1B（PTP1B）在unliganded（载脂蛋白）状态。在以前的载脂蛋白室温结构中，活性位点和变构位点采用交替构象，包括活性位点WPD环和远端变构位点的开放和闭合构象。相比之下，在我们的SSX结构中，活性位点最适合单一构象，但远端变构位点最适合替代构象。这一观察结果为解释这种蛋白质中变构偶联的性质提供了额外的细微差别。总的来说，我们的研究结果说明了室温晶体学系列方法的前景，以及未来的前卫晶体学实验，PTP1B和其他蛋白质。
The first serial synchrotron crystallographic structure of apo PTP1B is reported and the conformational heterogeneity at several allosteric sites is compared with that seen in prior structures. Room-temperature X-ray crystallography provides unique insights into protein conformational heterogeneity, but obtaining sufficiently large protein crystals is a common hurdle. Serial synchrotron crystallography (SSX) helps to address this hurdle by allowing the use of many medium- to small-sized crystals. Here, a recently introduced serial sample-support chip system has been used to obtain the first SSX structure of a human phosphatase, specifically protein tyrosine phosphatase 1B (PTP1B) in the unliganded (apo) state. In previous apo room-temperature structures, the active site and allosteric sites adopted alternate conformations, including open and closed conformations of the active-site WPD loop and of a distal allosteric site. By contrast, in our SSX structure the active site is best fitted with a single conformation, but the distal allosteric site is best fitted with alternate conformations. This observation argues for additional nuance in interpreting the nature of allosteric coupling in this protein. Overall, our results illustrate the promise of serial methods for room-temperature crystallography, as well as future avant-garde crystallography experiments, for PTP1B and other proteins.