Crystallization and preliminary X-ray analysis of cryptochrome 3 from Arabidopsis thaliana.
Cryptochromes are flavoproteins which serve as blue-light receptors in plants, animals, fungi and prokaryotes and belong to the same protein family as the catalytically active DNA photolyases. Cryptochrome 3 from the plant Arabidopsis thaliana (cry3; 525 amino acids, 60.7 kDa) is a representative of the novel cryDASH subfamily of UV-A/blue-light receptors and has been expressed as a mature FAD-containing protein in Escherichia coli without the signal sequence that directs the protein into plant organelles. The purified cryptochrome was found to be complexed to methenyltetrahydrofolate as an antenna pigment. Crystals of the cryptochrome-antenna pigment complex were obtained by vapour diffusion and display orthorhombic symmetry, with unit-cell parameters a = 76.298, b = 116.782, c = 135.024 A. X-ray diffraction data were collected to 1.9 A resolution using synchrotron radiation. The asymmetric unit comprises a cry3 dimer, the physiological role of which remains to be elucidated.
Pokorny
Richard
R
Philipps-Universität Marburg, FB Biologie, Karl-von-Frisch-Strasse 8, D-35032 Marburg, Germany.
Klar
Tobias
T
Essen
Lars-Oliver
LO
Batschauer
Alfred
A
D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins; D002384:Catalysis; D003001:Cloning, Molecular; D056931:Cryptochromes; D003460:Crystallization; D018360:Crystallography, X-Ray; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000737:chemistry; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry; D005420:Flavoproteins / Q000737:chemistry*; D008027:Light; D045102:Methylenetetrahydrofolate Dehydrogenase (NAD+) / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D013053:Spectrophotometry; D013329:Structure-Activity Relationship; D017356:Synchrotrons; D014466:Ultraviolet Rays; D014961:X-Ray Diffraction
D029681:Arabidopsis Proteins; C538962:CRY1 protein, Arabidopsis; D056931:Cryptochromes; D005420:Flavoproteins; D011994:Recombinant Proteins; D005182:Flavin-Adenine Dinucleotide; D045102:Methylenetetrahydrofolate Dehydrogenase (NAD+); D004255:Deoxyribodipyrimidine Photo-Lyase