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Rhodopsin pp 147–168Cite as

Crystallographic Studies of Rhodopsins: Structure and Dynamics

Part of the Methods in Molecular Biology book series (MIMB,volume 2501)

Abstract

Crystal structures have provided detailed insight in the architecture of rhodopsin photoreceptors. Of particular interest are the protein–chromophore interactions that govern the light-induced retinal isomerization and ultimately induce the large structural changes important for the various biological functions of the family. The reaction intermediates occurring along the rhodopsin photocycle have vastly differing lifetimes, from hundreds of femtoseconds to milliseconds. Detailed insight at high spatial and temporal resolution can be obtained by time-resolved crystallography using pump-probe approaches at X-ray free-electron lasers. Alternatively, cryotrapping approaches can be used. Both the approaches are described, including illumination and sample delivery. The importance of appropriate photoexcitation avoiding multiphoton absorption is stressed.

Key words

  • X-ray free-electron laser
  • XFEL
  • Serial femtosecond crystallography
  • SFX
  • Serial crystallography
  • Nanocrystallography
  • Time-resolved crystallography
  • Pump probe experiment
  • Photoexcitation
  • Rhodopsin
  • Retinal

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  • DOI: 10.1007/978-1-0716-2329-9_7
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Notes

  1. 1.

    The photocycle intermediates and their nomenclature in microbial rhodopsins were originally established in bR: K and O are red-shifted intermediates, L, M, and N are blue-shifted. Animal rhodopsins have similar intermediates called Batho, Lumi, Meta I, and Meta II. Additional transient species exist in both systems. The I (Photo) intermediate, an excited state, is formed within 200 fs, the K (Batho) intermediate within a few ps, Lumi in ns, L and then M (Meta I) in μs, N (Meta II) in ms, followed by O.

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Grünbein, M.L. et al. (2022). Crystallographic Studies of Rhodopsins: Structure and Dynamics. In: Gordeliy, V. (eds) Rhodopsin. Methods in Molecular Biology, vol 2501. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2329-9_7

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