A nanosecond laser spectroscopy platform for studying light-activated biomolecules

生物学中有许多过程是由光触发的（例如光合作用，感光蛋白，光激活酶或工程系统）。许多由暴露于光引起的这些变化发生在多个时间尺度上（通常从fs到s），并且涉及发色团和生物大分子（通常是蛋白质）中的化学和结构变化的复杂反应级联。这就需要对大范围时间尺度上的生物物理、光化学和结构变化进行比较分析。通过之前的BBSRC资助，我们已经定制并建立了一系列超快（亚纳秒）时间尺度的现场领先的时间分辨光谱仪器，具有可见光和红外吸收和荧光检测功能，以询问复杂的光驱动机制。虽然光敏生物过程需要在这些超快时间尺度上的激发态动力学，但驱动生物功能的光化学和结构变化通常发生在较慢的时间尺度上（纳秒-秒）。监测这些过程对于提供对机制的重要见解并为合理的重新设计提供信息至关重要。我们现在提出了一个多用途的时间分辨光谱仪器，以支持在英国生物科学界的广泛的计划。该仪器将具有广泛的能力，独特地包括在紫外可见光和红外区域的探测。拟议的技术平台包括一个纳秒激光系统，该系统将用于触发2个单独的时间分辨光谱仪，一个可以测量紫外-可见光区域的瞬时吸光度变化，另一个可以测量红外区域的瞬时吸光度变化，两者都在纳秒-毫秒的时间尺度上。这种新颖的实验装置，由真正的“即插即用”组件组成，将使我们能够将紫外可见光的变化与生物系统具有许多振动模式的中红外区域的变化相关联。这种仪器配置将是英国生物科学界所独有的，并将为曼彻斯特生物技术研究所的许多研究小组，更广泛的曼彻斯特大学和外部合作者提供重要的基础设施和技术资源，其中已经确定了一些研究项目。该综合设施将是一个以社区为基础的先进仪器，由专家实验科学家和国际公认的学术领导支持。

There are many processes in biology that are triggered by light (e.g. photosynthesis, photoreceptor proteins, light-activated enzymes or engineered systems). Many of these changes induced by exposure to light occur across multiple timescales (typically from fs - s), and involve complex reaction cascades of chemical and structural change in both chromophores and the biological macromolecule (typically protein). This requires comparative analysis of photophysical, photochemical and structural change across a wide range of timescale. Through previous BBSRC funding we have customised and built a range of field-leading time-resolved spectroscopy instruments on ultrafast (sub-nanosecond) timescales complete with visible and infra-red absorbance and fluorescence detection capabilities to interrogate complex light-driven mechanisms. Although light-sensitive biological processes require excited state dynamics on these ultrafast timescales the photochemical and structural changes that drive biological function generally occur on slower timescales (nanosecond-second). Monitoring these processes is crucial to provide important insight into mechanisms and informs rational re-design. We now propose a multi-purpose time-resolved spectroscopy instrument to support a wide range of programmes in the UK bioscience community. The instrumentation will have a wide range of capabilities that uniquely includes detection in both the UV-visible and infra-red regions. The proposed technology platform comprises a nanosecond laser system that will be used to trigger 2 separate time-resolved spectrometers, one that can measure transient absorbance changes in the UV-visible region and the other in the infra-red region, both on the nanosecond-millsecond timescale. This novel experimental set up, comprised of truly 'plug and play' components, will allow us to correlate changes in the UV-visible with those in the mid-infra-red region where biological systems have many vibrational modes. This instrumental configuration will be unique to the UK bioscience community and will provide significant infrastructure and technology resource for many research groups at the Manchester institute of Biotechnology, the wider University of Manchester and external collaborators, where a number of research projects have been identified. The integrated facility will be a community-based advanced instrument, supported by expert experimental scientists and internationally recognised academic leadership.