Suwa Yoshiaki, Nakamura Teruya, Toma Sachiko, Ikemizu Shinji, Kai Hirofumi, Yamagata Yuriko
Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Mar 1;64(Pt 3):171-4. doi: 10.1107/S1744309108002662. Epub 2008 Feb 23.
The Ets2 transcription factor is a member of the Ets transcription-factor family. Ets2 plays a role in the malignancy of cancer and in Down's syndrome by regulating the transcription of various genes. The DNA-binding domain of Ets2 (Ets domain; ETSD), which contains residues that are highly conserved among Ets transcription-factor family members, was expressed as a GST-fusion protein. The aggregation of ETSD produced after thrombin cleavage could be prevented by treatment with NDSB-195 (nondetergent sulfobetaine 195). ETSD was crystallized in complex with DNA containing the Ets2 target sequence (GGAA) by the hanging-drop vapour-diffusion method. The best crystals were grown using 25% PEG 3350, 80 mM magnesium acetate, 50 mM sodium cacodylate pH 5.0/5.5 as the reservoir at 293 K. The crystals belonged to space group C2, with unit-cell parameters a = 85.89, b = 95.52, c = 71.89 A, beta = 101.7 degrees and a V(M) value of 3.56 A(3) Da(-1). Diffraction data were collected to a resolution of 3.0 A.
Ets2转录因子是Ets转录因子家族的成员之一。Ets2通过调节各种基因的转录，在癌症的恶性发展和唐氏综合征中发挥作用。Ets2的DNA结合结构域（Ets结构域；ETSD）含有在Ets转录因子家族成员中高度保守的残基，被表达为一种GST融合蛋白。凝血酶切割后产生的ETSD聚集可通过用NDSB-195（非去污剂磺基甜菜碱195）处理来防止。通过悬滴气相扩散法，使ETSD与含有Ets2靶序列（GGAA）的DNA形成复合物并结晶。最佳晶体是在293 K下，以25% PEG 3350、80 mM乙酸镁、50 mM二甲胂酸钠pH 5.0/5.5作为储液生长得到的。晶体属于空间群C2，晶胞参数为a = 85.89、b = 95.52、c = 71.89 Å、β = 101.7°，V(M)值为3.56 ų Da⁻¹。收集到的衍射数据分辨率为3.0 Å。
