Dynamic photobiology: A collection of experimental protocols for time-resolved structural studies on light-activated...

The study of proteins that sense and respond to light reveals how nature has harnessed light for function while also mitigating its damaging effects through finely tuned molecular mechanisms. Time-resolved optical spectroscopy and time-resolved X-ray solution scattering and diffraction techniques are well-positioned to probe functional transitions triggered by light in photoactive proteins. However, integrating both fields presents technical and conceptual difficulties for many researchers. This FEBS Open Bio 'In the Limelight' special issue, Dynamic Photobiology, showcases five research protocols that provide comprehensive and practical guidance for designing and analyzing time-resolved experiments on proteins. The protocols cover studies performed in both solution and crystalline environments using time-resolved femtosecond Vis/IR spectroscopy, time-resolved X-ray solution scattering, and time-resolved serial crystallography at synchrotron and XFEL facilities. These detailed research protocols aim to ease collaboration across fields, acquaint early-career researchers with essential concepts, and offer a comprehensive overview for established researchers.