Origin of light-induced spin-correlated radical pairs in cryptochrome

J Phys Chem B. 2010 Nov 18;114(45):14745-54. doi: 10.1021/jp103401u. Epub 2010 Aug 4.

Abstract

Blue-light excitation of cryptochromes and homologues uniformly triggers electron transfer (ET) from the protein surface to the flavin adenine dinucleotide (FAD) cofactor. A cascade of three conserved tryptophan residues has been considered to be critically involved in this photoreaction. If the FAD is initially in its fully oxidized (diamagnetic) redox state, light-induced ET via the tryptophan triad generates a series of short-lived spin-correlated radical pairs comprising an FAD radical and a tryptophan radical. Coupled doublet-pair species of this type have been proposed as the basis, for example, of a biological magnetic compass in migratory birds, and were found critical for some cryptochrome functions in vivo. In this contribution, a cryptochrome-like protein (CRYD) derived from Xenopus laevis has been examined as a representative system. The terminal radical-pair state FAD(•)···W324(•) of X. laevis CRYD has been characterized in detail by time-resolved electron-paramagnetic resonance (TREPR) at X-band microwave frequency (9.68 GHz) and magnetic fields around 345 mT, and at Q-band (34.08 GHz) at around 1215 mT. Different precursor states, singlet versus triplet, of radical-pair formation have been considered in spectral simulations of the experimental electron-spin polarized TREPR signals. Conclusively, we present evidence for a singlet-state precursor of FAD(•)···W324(•) radical-pair generation because at both magnetic fields, where radical pairs were studied by TREPR, net-zero electron-spin polarization has been detected. Neither a spin-polarized triplet precursor nor a triplet at thermal equilibrium can explain such an electron-spin polarization. It turns out that a two-microwave-frequency TREPR approach is essential to draw conclusions on the nature of the precursor electronic states in light-induced spin-correlated radical pair formations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cryptochromes / chemistry*
  • Cryptochromes / metabolism*
  • Electron Spin Resonance Spectroscopy
  • Electron Transport / radiation effects
  • Flavin-Adenine Dinucleotide / metabolism
  • Free Radicals / chemistry
  • Free Radicals / metabolism
  • Light*
  • Models, Molecular
  • Protein Conformation
  • Xenopus Proteins / chemistry
  • Xenopus Proteins / metabolism
  • Xenopus laevis

Substances

  • Cryptochromes
  • Free Radicals
  • Xenopus Proteins
  • Flavin-Adenine Dinucleotide