TY  - JOUR
AU  - Kottke, T.
AU  - Ahmad, M.
AU  - Batschauer, A.
AU  - Heberle, J.
TI  - Blue-Light-Induced Changes in Arabidopsis Cryptochrome 1 Probed by FTIR Difference Spectroscopy
JO  - Biochemistry
VL  - 45
SN  - 0006-2960
CY  - Columbus, Ohio
PB  - American Chemical Society
M1  - PreJuSER-51338
SP  - 2472 - 2479
PY  - 2006
N1  - Record converted from VDB: 12.11.2012
AB  - Cryptochromes are blue-light photoreceptors that regulate a variety of responses in animals and plants, including circadian entrainment in Drosophila and photomorphogenesis in Arabidopsis. They comprise a photolyase homology region (PHR) of about 500 amino acids and a C-terminal extension of varying length. In the PHR domain, flavin adenine dinucleotide (FAD) is noncovalently bound. The presence of a second chromophore, such as methenyltetrahydrofolate, in animal and plant cryptochromes is still under debate. Arabidopsis cryptochrome 1 (CRY1) has been intensively studied with regard to function and interaction of the protein in vivo and in vitro. However, little is known about the pathway from light absorption to signal transduction on the molecular level. We investigated the full-length CRY1 protein by Fourier transform infrared (FTIR) and UV/vis difference spectroscopy. Starting from the fully oxidized state of the chromophore FAD, a neutral flavoprotein radical is formed upon illumination in the absence of any exogenous electron donor. A preliminary assignment of the chromophore bands is presented. The FTIR difference spectrum reveals only moderate changes in secondary structure of the apoprotein in response to the photoreduction of the chromophore. Deprotonation of an aspartic or glutamic acid, probably D396, accompanies radical formation, as is deduced from the negative band at 1734 cm(-)(1) in D(2)O. The main positive band at 1524 cm(-)(1) in the FTIR spectrum shows a strong shift to lower frequencies as compared to other flavoproteins. Together with the unusual blue-shift of the absorption in the visible range to 595 nm, this clearly distinguishes the radical form of CRY1 from those of structurally highly homologous DNA photolyases. As a consequence, the direct comparison of cryptochrome to photolyase in terms of photoreactivity and mechanism has to be made with caution.
KW  - Arabidopsis Proteins: chemistry
KW  - Cryptochromes
KW  - Crystallography, X-Ray
KW  - Deoxyribodipyrimidine Photo-Lyase: chemistry
KW  - Deoxyribodipyrimidine Photo-Lyase: metabolism
KW  - Escherichia coli: metabolism
KW  - Flavins: metabolism
KW  - Flavoproteins: chemistry
KW  - Flavoproteins: metabolism
KW  - Light
KW  - Models, Molecular
KW  - Signal Transduction: physiology
KW  - Spectrophotometry, Ultraviolet
KW  - Spectroscopy, Fourier Transform Infrared: methods
KW  - Structural Homology, Protein
KW  - Arabidopsis Proteins (NLM Chemicals)
KW  - Cryptochromes (NLM Chemicals)
KW  - Flavins (NLM Chemicals)
KW  - Flavoproteins (NLM Chemicals)
KW  - Deoxyribodipyrimidine Photo-Lyase (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:16489739
UR  - <Go to ISI:>//WOS:000235792300002
DO  - DOI:10.1021/bi051964b
UR  - https://juser.fz-juelich.de/record/51338
ER  -