Serial Femtosecond Crystallography of Optogenetic Function

光发生功能的系列飞秒晶体学

基本信息

批准号：
BB/P00752X/1
负责人：
Jasper Van Thor
金额：
$ 82.73万
依托单位：
Imperial College London
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2017
资助国家：
英国
起止时间：
2017 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FP00752X%2F1
关键词：
Serial Femtosecond Crystallography Optogenetic Function

项目摘要

The ability to design synthetic light-sensitive materials that can be genetically encoded provides biologists the means and opportunity to sense and control biological function and environment. In order to learn how these materials function it has very recently become possible to make very fast 'snapshots' of the light-induced motions using X-ray crystallography, which is even possible for very short time scales that still involves the excited electrons (on femtosecond time scale). Novel 4th generation light sources can now be used to record such 'molecular movies' which is a breakthrough technology that we only very recently demonstrated. In the very near future we anticipate being able to record even better quality movies, as the European XFEL in Hamburg that allows much more data to be recorded will start user operation. Where previously we could only use laser spectroscopy techniques we can now actually 'watch' the very first motions of proteins directly after activation. It is known that the 'outcome' of biological reactions involving either sensing or activation (via 'actuation') is determined in these very first motions that occur on time scales typically less than picoseconds. For example, in the case of a bi-directional photochromic fluorescent protein the efficiency of switching in one direction is excellent at 30%, while the efficiency of switching in the reverse direction is poor at less than 0.05%, potentially limiting optogenetics applications. We are thus in a position to ask the question what determines this dramatic difference in the outcome of the reactions, and provide structural and dynamical information that will be highly valuable for feeding back for rational optogenetics design.

设计可进行基因编码的合成光敏材料的能力为生物学家提供了感知和控制生物功能和环境的手段和机会。为了了解这些材料的功能，最近可以使用 X 射线晶体学对光诱导运动进行非常快速的“快照”，这甚至可以在仍然涉及激发电子的非常短的时间尺度内进行（在飞秒时间尺度）。现在可以使用新型第四代光源来记录此类“分子电影”，这是我们最近才展示的一项突破性技术。在不久的将来，我们预计能够录制质量更高的电影，因为位于汉堡的欧洲 XFEL 可以录制更多数据，并将开始用户操作。以前我们只能使用激光光谱技术，现在我们实际上可以直接“观察”蛋白质激活后的第一个运动。众所周知，涉及传感或激活（通过“驱动”）的生物反应的“结果”是在这些最初的运动中确定的，这些运动的时间尺度通常小于皮秒。例如，在双向光致变色荧光蛋白的情况下，一个方向的转换效率高达 30%，而相反方向的转换效率较差，低于 0.05%，这可能限制了光遗传学的应用。因此，我们能够提出一个问题，是什么决定了反应结果的这种巨大差异，并提供结构和动力学信息，这些信息对于反馈合理的光遗传学设计非常有价值。