Photoactivation of the flavin cofactor in Xenopus laevis (6–4) photolyase: Observation of a transient tyrosyl radical by time-resolved electron paramagnetic resonance

The light-induced electron transfer reaction of flavin cofactor photoactivation in Xenopus laevis (6–4) photolyase has been studied by continuous-wave and time-resolved electron paramagnetic resonance spectroscopy. When the photoactivation is initiated from the fully oxidized form of the flavin, a neutral flavin radical is observed as a long-lived paramagnetic intermediate of two consecutive single-electron reductions under participation of redox-active amino acid residues. By time-resolved electron paramagnetic resonance, a spin-polarized transient radical-pair signal was detected that shows remarkable differences to the signals observed in the related cyclobutane pyrimidine dimer photolyase enzyme. In (6–4) photolyase, a neutral tyrosine radical has been identified as the final electron donor, on the basis of the characteristic line width, hyperfine splitting pattern, and resonance magnetic field position of the tyrosine resonances of the transient radical pair.

[1]  W. Lubitz,et al.  Tryptophan and tyrosine radicals in ribonucleotide reductase: a comparative high-field EPR study at 94 GHz. , 2001, Biochemistry.

[2]  T. Carell,et al.  The mechanism of action of DNA photolyases. , 2001, Current opinion in chemical biology.

[3]  J. Deisenhofer.  DNA photolyases and cryptochromes. , 2000, Mutation research.

[4]  G. Sancar,et al.  Enzymatic photoreactivation: 50 years and counting. , 2000, Mutation research.

[5]  A. Stuchebrukhov,et al.  Calculation of Quantum Parameters for Nonadiabatic Redox Reactions. Application to Photoreduction of Flavin in DNA Photolyase , 2000 .

[6]  A. Eker,et al.  Intraprotein radical transfer during photoactivation of DNA photolyase , 2000, Nature.

[7]  A. Bacher,et al.  EPR, ENDOR, and TRIPLE resonance spectroscopy on the neutral flavin radical in Escherichia coli DNA photolyase. , 1999, Biochemistry.

[8]  A. Boussac,et al.  EPR Detection of the Transient Tyrosyl Radical in DNA Photolyase from Anacystis nidulans , 1999 .

[9]  T. Todo,et al.  Functional diversity of the DNA photolyase/blue light receptor family. , 1999, Mutation research.

[10]  A. Eker,et al.  Intraprotein electron transfer between tyrosine and tryptophan in DNA photolyase from Anacystis nidulans. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Cashmore,et al.  Cryptochromes: blue light receptors for plants and animals. , 1999, Science.

[12]  A. Sancar,et al.  Origin of the Transient Electron Paramagnetic Resonance Signals in DNA Photolyase , 1999 .

[13]  A. Stuchebrukhov,et al.  Pathways of electron transfer in Escherichia coli DNA photolyase: Trp306 to FADH. , 1999, Biophysical journal.

[14]  D. Mu,et al.  (6-4) photolyase: light-dependent repair of DNA damage. , 1998, Histology and histopathology.

[15]  T. Todo,et al.  Binding and Catalytic Properties of Xenopus (6-4) Photolyase* , 1997, The Journal of Biological Chemistry.

[16]  D. S. Hsu,et al.  Reaction Mechanism of (6-4) Photolyase* , 1997, The Journal of Biological Chemistry.

[17]  L. Eriksson,et al.  Theoretical Study of Model Tryptophan Radicals and Radical Cations: Comparison with Experimental Data of DNA Photolyase, Cytochrome c Peroxidase, and Ribonucleotide Reductase , 1997 .

[18]  Satoru Kanai,et al.  Molecular Evolution of the Photolyase–Blue-Light Photoreceptor Family , 1997, Journal of Molecular Evolution.

[19]  F. Himo,et al.  Density functional calculations on model tyrosyl radicals. , 1997, Biophysical journal.

[20]  T. Todo,et al.  Flavin adenine dinucleotide as a chromophore of the Xenopus (6-4)photolyase. , 1997, Nucleic acids research.

[21]  C. Gray,et al.  Chattering Cells: Superficial Pyramidal Neurons Contributing to the Generation of Synchronous Oscillations in the Visual Cortex , 1996, Science.

[22]  T. Todo,et al.  Similarity Among the Drosophila (6-4)Photolyase, a Human Photolyase Homolog, and the DNA Photolyase-Blue-Light Photoreceptor Family , 1996, Science.

[23]  B. Diner,et al.  245 GHz High-Field EPR Study of Tyrosine-D° and Tyrosine-Z° in Mutants of Photosystem II† , 1996 .

[24]  J. Deisenhofer,et al.  Crystal structure of DNA photolyase from Escherichia coli. , 1995, Science.

[25]  R. Rustandi,et al.  Photoinduced spin-polarized radical pair formation in a DNA photolyase.substrate complex at low temperature. , 1995, Biochemistry.

[26]  J. Norris,et al.  Transient EPR of light-induced spin-correlated radical pairs. Manifestation of zero quantum coherence , 1994 .

[27]  A. Sancar.  Structure and function of DNA photolyase. , 1994, Biochemistry.

[28]  A. Sancar,et al.  Time-resolved EPR studies with DNA photolyase: excited-state FADH0 abstracts an electron from Trp-306 to generate FADH-, the catalytically active form of the cofactor. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[29]  T. Todo,et al.  A new photoreactivating enzyme that specifically repairs ultraviolet light-induced (6-4)photoproducts , 1993, Nature.

[30]  A. Sancar,et al.  Active site of DNA photolyase: tryptophan-306 is the intrinsic hydrogen atom donor essential for flavin radical photoreduction and DNA repair in vitro. , 1991, Biochemistry.

[31]  A. Sancar,et al.  Excited-state properties of Escherichia coli DNA photolyase in the picosecond to millisecond time scale. , 1990, Biochemistry.

[32]  C. Walsh,et al.  Purification and properties of Methanobacterium thermoautotrophicum DNA photolyase. , 1989, The Journal of biological chemistry.

[33]  J. Petersen,et al.  Anisotropic electron spin polarization of correlated spin pairs in photosynthetic reaction centers , 1989 .

[34]  J. D. Velde,et al.  Photoreactivating enzyme from the green alga Scenedesmus acutus. Evidence for the presence of two different flavin chromophores , 1988 .

[35]  A. Sancar,et al.  The active form of Escherichia coli DNA photolyase contains a fully reduced flavin and not a flavin radical, both in vivo and in vitro. , 1987, Biochemistry.

[36]  D. A. Hunter,et al.  Electron paramagnetic resonance of spin-correlated radical pairs in photosynthetic reactions , 1987 .

[37]  J. Norris,et al.  Spin-polarized electron paramagnetic resonance spectra of radical pairs in micelles: observation of electron spin-spin interactions , 1987 .

[38]  A. Sancar,et al.  Action mechanism of Escherichia coli DNA photolyase. III. Photolysis of the enzyme-substrate complex and the absolute action spectrum. , 1987, The Journal of biological chemistry.

[39]  A. Sancar,et al.  Photochemical properties of Escherichia coli DNA photolyase: a flash photolysis study. , 1986, Biochemistry.

[40]  A. Sancar,et al.  Identification of a neutral flavin radical and characterization of a second chromophore in Escherichia coli DNA photolyase. , 1984, Biochemistry.

[41]  A. Sancar,et al.  Escherichia coli DNA photolyase is a flavoprotein. , 1984, Journal of molecular biology.

[42]  V. Massey,et al.  On the existence of spectrally distinct classes of flavoprotein semiquinones. A new method for the quantitative production of flavoprotein semiquinones. , 1966, Biochemistry.

[43]  H. Beinert.  Spectral Characteristics of Flavins at the Semiquinoid Oxidation Level1 , 1956 .

[44]  T. Todo,et al.  Cloning and characterization of a gene (UVR3) required for photorepair of 6-4 photoproducts in Arabidopsis thaliana. , 1998, Nucleic acids research.

[45]  F. Müller.  The flavin redox-system and its biological function. , 1983, Topics in current chemistry.

[46]  W. T. Dixon,et al.  Determination of the acidity constants of some phenol radical cations by means of electron spin resonance , 1976 .