Crystallization and preliminary X-ray analysis of cryptochrome 3 from Arabidopsis thaliana.
Crystallization and preliminary X-ray analysis of cryptochrome 3 from Arabidopsis thaliana.
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DOI：
10.1107/s1744309105028897
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发表时间：
2005-10
期刊：
影响因子：
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通讯作者：
R. Pokorný;T. Klar;L. Essen;A. Batschauer
中科院分区：
文献类型：
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作者：
R. Pokorný;T. Klar;L. Essen;A. Batschauer
隐花色素是植物、动物、真菌和原核生物中作为蓝光受体的黄素蛋白，与具有催化活性的DNA光解酶属于同一蛋白家族。来自植物拟南芥的隐花色素3（cry 3; 525个氨基酸，60.7kDa）是UV-A/蓝光受体的新型cryDASH亚家族的代表，并且已经在大肠杆菌中表达为成熟的含FAD的蛋白质，而没有指导蛋白质进入植物细胞器的信号序列。纯化的隐花色素被发现是复杂的亚甲基四氢叶酸作为天线色素。用气相扩散法获得了隐花色素-触角色素配合物晶体，晶体具有正交对称性，晶胞参数a = 76.298，B = 116.782，c = 135.024 A。使用同步辐射收集X射线衍射数据至1.9 A分辨率。不对称单元包括cry 3二聚体，其生理作用仍有待阐明。
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.