C-Terminal Extension of a Plant Cryptochrome Dissociates from the β-Sheet of the Flavin-Binding Domain

J Phys Chem Lett. 2021 Jun 17;12(23):5558-5563. doi: 10.1021/acs.jpclett.1c00844. Epub 2021 Jun 8.

Abstract

Plant cryptochromes are central blue light receptors in land plants and algae. Photoreduction of the flavin bound to the photolyase homology region (PHR) causes a dissociation of the C-terminal extension (CCT) as effector via an unclear pathway. We applied the recently developed in-cell infrared difference (ICIRD) spectroscopy to study the response of the full-length pCRY from Chlamydomonas reinhardtii in living bacterial cells, because the receptor degraded upon isolation. We demonstrate a stabilization of the flavin neutral radical as photoproduct and of the resulting β-sheet reorganization by binding of cellular ATP. Comparison between light-induced structural responses of full-length pCRY and PHR reveals a downshift in frequency of the β-sheet signal, implying an association of the CCT close to the only β-sheet of the PHR in the dark. We provide a missing link in activation of plant cryptochromes after flavin photoreduction by indicating that β-sheet reorganization causes the CCT release and restructuring.

MeSH terms

  • Binding Sites / physiology
  • Chlamydomonas reinhardtii / chemistry*
  • Chlamydomonas reinhardtii / metabolism*
  • Cryptochromes / analysis
  • Cryptochromes / chemistry*
  • Cryptochromes / metabolism*
  • Flavins / analysis
  • Flavins / chemistry*
  • Flavins / metabolism*
  • Protein Conformation, beta-Strand
  • Protein Structure, Secondary
  • Spectrophotometry, Infrared / methods

Substances

  • Cryptochromes
  • Flavins