A nanosecond laser spectroscopy platform for studying light-activated biomolecules

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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.