Purification, crystallization and preliminary X-ray data collection of the N-terminal domain of the 26S proteasome regulatory subunit p27 and its complex with the ATPase domain of Rpt5 from Mus musculus
Purification, crystallization and preliminary X-ray data collection of the N-terminal domain of the 26S proteasome regulatory subunit p27 and its complex with the ATPase domain of Rpt5 from Mus musculus
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小家鼠 26S 蛋白酶体调节亚基 p27 的 N 端结构域及其与 Rpt5 ATPase 结构域的复合物的纯化、结晶和初步 X 射线数据收集
DOI：
10.1107/s2053230x14006815
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发表时间：
2014-05-01
影响因子：
0.9
通讯作者：
Zhou, Hao
中科院分区：
文献类型：
--
作者：
Diao, Wentao;Yang, Xue;Zhou, Hao
蛋白质26 S蛋白酶体调节亚基p27是帮助26 S蛋白酶体的19 S调节颗粒（RP）组装的四种分子伴侣之一。本研究克隆、表达、纯化并结晶了小家鼠p27的N端（残基1-128），以及与Rpt 5的C端ATP酶结构域（残基173-442）复合的蛋白。p27（1-128）晶体的衍射分辨率为1.7埃，空间群为P2（1）2（1）2（1），晶胞参数a = 26.79，B = 30.39，c = 145.06。分辨率依赖的马修斯系数概率分析表明，每个不对称单元仅存在一个分子，溶剂含量为40.5%，V-M值为2.02埃（3）Da（-1）。p27（（1-128））-Rpt 5（（173-442））复合物的晶体衍射分辨率为4埃，属于空间群P222，晶胞参数a = 75.93，B = 76.08，c = 336.85埃。假设在具有53.2%溶剂含量和2.63埃（3）Da（-1）的V-M值的不对称单元中存在四种异二聚体，或者在具有41.5%溶剂含量和2.10埃（3）Da（-1）的V-M值的不对称单元中存在五种异二聚体。
The protein 26S proteasome regulatory subunit p27 is one of the four chaperones that help in the assembly of the 19S regulatory particle (RP) of the 26S proteasome. In the present work, the N-terminus of p27 (residues 1-128) from Mus musculus was cloned, expressed, purified and crystallized alone and in complex with the C-terminal ATPase domain of Rpt5 (residues 173-442). The crystals of p27 (1-128) diffracted to 1.7 angstrom resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 26.79, b = 30.39, c = 145.06 A. Resolution-dependent Matthews coefficient probability analysis suggested the presence of only one molecule per asymmetric unit, with 40.5% solvent content and a V-M value of 2.02 angstrom(3) Da(-1). The crystal of the p27((1-128))-Rpt5((173-442)) complex diffracted to 4 angstrom resolution and belonged to space group P222, with unit-cell parameters a = 75.93, b = 76.08, c = 336.85 angstrom. The presence of four heterodimers in the asymmetric unit with 53.2% solvent content and a V-M value of 2.63 angstrom(3) Da(-1) or five heterodimers in the asymmetric unit with 41.5% solvent content and a V-M value of 2.10 angstrom(3) Da(-1) is assumed.