A guide to time-resolved structural analysis of light-activated proteins

Harshwardhan Poddar, Derren Heyes, Giorgio Schirò, Martin Weik , David Leys, Nigel Scrutton

Research output: Contribution to journalArticlepeer-review


Dynamical changes in protein structures are essential for protein function and occur over femtoseconds to seconds timescales. X-ray free electron lasers have facilitated investigations of structural dynamics in proteins with unprecedented temporal and spatial resolution. Light-activated proteins are attractive targets for time-resolved structural studies, as the reaction chemistry and associated protein structural changes can be triggered by short laser pulses. Proteins with different light-absorbing centres have evolved to detect light and harness photon energy to bring about downstream chemical and biological output responses. Following light absorption, rapid chemical/small-scale structural changes are typically localised around the chromophore. These localised changes are followed by larger structural changes propagated throughout the photoreceptor/photocatalyst that enables the desired chemical and/or biological output response. Time-resolved serial femtosecond crystallography (SFX) and solution scattering techniques enable direct visualisation of early chemical change in light-activated proteins on timescales previously inaccessible, whereas scattering gives access to slower timescales associated with more global structural change. Here, we review how advances in time-resolved SFX and solution scattering techniques have uncovered mechanisms of photochemistry and its coupling to output responses. We also provide a prospective on how these time-resolved structural approaches might impact on other photoreceptors/photoenzymes that have not yet been studied by these methods.
Original languageEnglish
Pages (from-to)576-595
Number of pages20
JournalThe FEBS Journal
Issue number3
Early online date17 Apr 2021
Publication statusPublished - 1 Feb 2022


  • light-activated proteins
  • photoenzymes
  • photoreceptors
  • protein dynamics
  • SAXS
  • time-resolved
  • TR-SFX
  • WAXS

Research Beacons, Institutes and Platforms

  • Manchester Institute of Biotechnology


Dive into the research topics of 'A guide to time-resolved structural analysis of light-activated proteins'. Together they form a unique fingerprint.

Cite this