Crystallization and preliminary crystallographic analysis of a PHD domain of human JARID1B
Crystallization and preliminary crystallographic analysis of a PHD domain of human JARID1B
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人 JARID1B PHD 结构域的结晶和初步晶体学分析
DOI：
10.1107/s1744309111021981
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发表时间：
2011-08-01
影响因子：
0.9
通讯作者：
Yang, Huirong
中科院分区：
文献类型：
--
作者：
Guo, Xue;Xu, Youwei;Yang, Huirong
含有 JmjC 结构域的蛋白质可以以序列和甲基化状态特异性的方式去除组蛋白赖氨酸甲基化。 JARID1B 是一种包含 PHD 和 JmjC 结构域的蛋白质，是 H3K4me2 和 H3K4me3 特异性的组蛋白去甲基酶，需要 Fe(II) 和 α-酮戊二酸 (α-KG) 作为辅助因子来去除甲基。 JARID1B 也被证明在乳腺癌的发展中发挥着关键作用。 JARID1B 包含 JmjN、Arid 和 JmjC 结构域、一个 C5HC2 锌指结构域和三个 PHD 结构域。第一个 PHD 结构域（PHD1(JARID1B)；残基 306-360）位于 N 末端，对于组蛋白去甲基化酶活性和 JARID1B 的组蛋白尾识别都很重要。在此，报道了 PHD1(JARID1B) 的表达、纯化和结晶。 PHD1(JARID1B) 晶体通过悬滴蒸气扩散法在由 0.1 M HEPES pH 7.0、2.2 M 硫酸铵组成的储库溶液中在 277 K 下生长。从 PHD1(JARID1B) 晶体收集锌 SAD 数据集。使用同步加速器辐射，PHD1(JARID1B) 晶体的衍射图样扩展至 1.65 埃分辨率。该晶体属于P4(3)空间群，晶胞参数a = 51.7，b = 51.7，c = 36.2埃。
Histone lysine methylation can be removed by proteins containing JmjC domains in a sequence- and methylation state-specific manner. JARID1B, a protein containing PHD and JmjC domains, is a histone demethylase specific for H3K4me2 and H3K4me3 which requires Fe(II) and alpha-ketoglutarate (alpha-KG) as cofactors to remove the methyl group. JARID1B has also been shown to play a critical role in the development of breast cancer. JARID1B contains JmjN, Arid and JmjC domains, a C5HC2 zinc-finger domain and three PHD domains. The first PHD domain (PHD1(JARID1B); residues 306-360) is located at the N-terminus and is important for both histone demethylase activity and histone-tail recognition of JARID1B. Here, the expression, purification and crystallization of PHD1(JARID1B) is reported. A PHD1(JARID1B) crystal was grown by the hanging-drop vapour-diffusion method in reservoir solution consisting of 0.1 M HEPES pH 7.0, 2.2 M ammonium sulfate at 277 K. A zinc SAD data set was collected from a PHD1(JARID1B) crystal. The diffraction pattern of the PHD1(JARID1B) crystal extended to 1.65 angstrom resolution using synchrotron radiation. The crystal belonged to space group P4(3), with unit-cell parameters a = 51.7, b = 51.7, c = 36.2 angstrom.