Purification, crystallization and preliminary X-ray diffraction analysis of the Kelch-like motif region of mouse Keap1
Purification, crystallization and preliminary X-ray diffraction analysis of the Kelch-like motif region of mouse Keap1
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DOI：
10.1107/s1744309104032506
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发表时间：
2005-01-01
影响因子：
0.9
通讯作者：
Yokoyama, S
中科院分区：
文献类型：
--
作者：
Padmanabhan, B;Scharlock, M;Yokoyama, S
Keap 1（Kelch-like ECH-associating protein 1）是细胞质中Nrf 2转录因子的负调节因子。Keap 1的Kelch/DGR（双甘氨酸重复）结构域与Nrf 2以及肌动蛋白丝相关。在大肠杆菌中表达含有小鼠Keap 1的Kelch/DGR结构域和C-末端区域的重组蛋白，通过坐滴气相扩散法纯化至接近均一并结晶。晶体属P6（1）或P6（5）空间群，晶胞参数a = B = 102.95，c = 55.21埃，不对称单元中含有一个分子。使用安装在RA-Micro 7 Cu K α双阳极X射线发生器上的R-AXIS IV++成像板收集完整的衍射数据至2.25埃分辨率。
Keap1 (Kelch-like ECH-associating protein 1) is a negative regulator of the Nrf2 transcription factor in the cytoplasm. The Kelch/DGR (double-glycine repeat) domain of Keap1 associates with Nrf2 as well as with actin filaments. A recombinant protein containing both the Kelch/DGR domain and the C-terminal region of mouse Keap1 was expressed in Escherichia coli, purified to near-homogeneity and crystallized by the sitting-drop vapour-diffusion method. The crystal belongs to space group P6(1) or P6(5), with unit-cell parameters a = b = 102.95, c = 55.21 angstrom, and contains one molecule in the asymmetric unit. A complete diffraction data was collected to 2.25 angstrom resolution using an R-AXIS IV++ imaging plate mounted on an RA-Micro7 Cu K alpha rotating-anode X-ray generator.