Characterization and crystallization of human DPY-30-like protein, an essential component of dosage compensation complex
Characterization and crystallization of human DPY-30-like protein, an essential component of dosage compensation complex
复制标题
DOI：
10.1016/j.bbapap.2005.08.011
复制
发表时间：
2005-12-01
影响因子：
3.2
通讯作者：
Rao, ZH
中科院分区：
文献类型：
--
作者：
Dong, XH;Peng, Y;Rao, ZH
人DPY-30样是秀丽线虫DPY-30的同源物。DPY-30是剂量补偿机制的重要组成部分，DPY-30活性的丧失导致XX特异性致死。在XO动物中，DPY-30是发育过程所必需的，而不是剂量补偿。在酵母中，DPY-30的同源物Saf 19 p作为组蛋白3赖氨酸4甲基化复合物的成员发挥功能，是表观遗传发育控制的关键部分。在这份报告中，人DPY-30样蛋白的过表达和纯化的结构确定的目标。在291 K下，通过气相扩散技术从由（NH_4）（2）SO_4（1.5-2.0 M）、Tris-HCl（0.1 M，pH 8.0）组成的沉淀剂溶液中以悬滴形式结晶。该晶体在100 K下内部衍射至2.7埃分辨率，属于空间群P4（1）2（1）2或P4（3）2（1）2，晶胞参数为a=B = 74.5埃，c = 87.0埃，α = β = γ =90.0度。不对称单元包含两个分子，溶剂含量为49%。我们还分析了它的生物化学和生物物理特性。目前正在努力确定DPY-30样的分子结构。这些研究将为基于结构的人DPY-30样和剂量补偿机制的功能分析开辟新的途径。(c)2005年由Elsevier B. V.出版
Human DPY-30-like is a homolog of C elegans DPY-30. DPY-30 is an essential component of dosage compensation machinery and loss of dpy-30 activity results in XX-specific lethality. In XO animals, DPY-30 is required for developmental processes other than dosage compensation. In yeast, the homolog of DPY-30, Saf19p, functions as a member of histone 3 lysine 4 methylation complex, which is the key part of epigenetic developmental control. In this report, human DPY-30-like protein was overexpressed and purified with the goal of structure determination. It was crystallized at 291 K in hanging drops by the vapour diffusion technique from a precipitant solution consisting of (NH4)(2)SO4 (1.5-2.0 M), Tris-HCl (0.1 M, pH 8.0). The crystal diffracted to 2.7 angstrom resolution at 100 K in-house and belongs to the space group P4(1)2(1)2 or P4(3)2(1)2 with unit-cell parameters of a=b = 74.5 angstrom, c = 87.0 angstrom, alpha = beta = gamma =90.0 degrees. The asymmetric unit contains two molecules with 49% solvent content. We also analyzed its biochemical and biophysical characterizations. Efforts are now under way to determine the molecular structure of the DPY-30-like. These studies will open a new avenue towards the structure-based functional analysis of human DPY-30-like and dosage compensation machinery. (c) 2005 Published by Elsevier B.V.