Crystallization and preliminary X-ray diffraction analysis of YidC, a membrane-protein chaperone and insertase from Bacillus halodurans.
Crystallization and preliminary X-ray diffraction analysis of YidC, a membrane-protein chaperone and insertase from Bacillus halodurans.
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DOI：
10.1107/s2053230x14012540
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发表时间：
2014-08
期刊：
影响因子：
--
通讯作者：
Nureki O
中科院分区：
文献类型：
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作者：
Kumazaki K;Tsukazaki T;Nishizawa T;Tanaka Y;Kato HE;Nakada-Nakura Y;Hirata K;Mori Y;Suga H;Dohmae N;Ishitani R;Nureki O
YidC，来自B的膜蛋白伴侣/插入酶。halodurans，表达，纯化和结晶在β-dic立方相中。收集X射线衍射数据集至2.4 μ m分辨率。 YidC是YidC/Oxa 1/Alb 3家族的成员，它将蛋白质插入细胞膜，并促进细菌中的膜蛋白折叠。YidC在Sec介导的整合和Sec非依赖性的膜蛋白插入中起关键作用。在这里，耐盐芽孢杆菌YidC 2，其中有五个跨膜螺旋之间的其他家庭成员的保守，被确定为目标蛋白的结构测定的荧光尺寸排阻色谱分析。该蛋白质被过表达，纯化和结晶在无规立方相中。晶体衍射X射线分辨率为2.4 Å，属于空间群P21，晶胞参数a = 43.9，B = 60.6，c = 58.9 Å，β = 100.3°。 实验阶段确定的多波长异常衍射方法，使用汞衍生的晶体。
YidC, a membrane-protein chaperone/insertase from B. halodurans, was expressed, purified and crystallized in the lipidic cubic phase. An X-ray diffraction data set was collected to 2.4 Å resolution. YidC, a member of the YidC/Oxa1/Alb3 family, inserts proteins into the membrane and facilitates membrane-protein folding in bacteria. YidC plays key roles in both Sec-mediated integration and Sec-independent insertion of membrane proteins. Here, Bacillus halodurans YidC2, which has five transmembrane helices conserved among the other family members, was identified as a target protein for structure determination by a fluorescent size-exclusion chromatography analysis. The protein was overexpressed, purified and crystallized in the lipidic cubic phase. The crystals diffracted X-rays to 2.4 Å resolution and belonged to space group P21, with unit-cell parameters a = 43.9, b = 60.6, c = 58.9 Å, β = 100.3°. The experimental phases were determined by the multiwavelength anomalous diffraction method using a mercury-derivatized crystal.