Projection structure of P-glycoprotein by electron microscopy - Evidence for a closed conformation of the nucleotide binding domains
Projection structure of P-glycoprotein by electron microscopy - Evidence for a closed conformation of the nucleotide binding domains
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DOI：
10.1074/jbc.m206871200
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发表时间：
2002-10-18
影响因子：
4.8
通讯作者：
Wilkens, S
中科院分区：
文献类型：
--
作者：
Lee, JY;Urbatsch, IL;Wilkens, S
用电子显微镜和图像分析技术对小鼠P-糖蛋白的结构进行了研究。利用脂质单层技术，通过重组含有C-末端六个组氨酸标签的纯净的洗涤剂溶解的蛋白质，在脂质双层中产生了PGP的二维晶体。晶体属于平面群P1，a=b=104+/-2埃，伽马=90+/-4度。在22A分辨率下计算的PGP的投影结构显示了两个紧密相互作用的蛋白质结构域，可以解释为蛋白质的N-末端和C-末端的一半。Pgp的投影结构与最近发表的MSBA的X射线结构一致，MSBA是一种来自大肠杆菌的脂类A翻转酶，与Pgp具有很高的序列同源性，但只有当两个MSBA亚基旋转时才能将它们的核苷酸结合域结合在一起。
The structure of P-glycoprotein (Pgp) from mouse has been studied by electron microscopy and image analysis. Two-dimensional crystals of Pgp in a lipid bilayer were generated by reconstituting pure, detergent-solubilized protein containing a C-terminal six-histidine tag using the lipid monolayer technique. The crystals belong to plane group P1 with a = b = 104 +/- 2 Angstrom and gamma = 90 +/- 4degrees. The projection structure of Pgp calculated at a resolution of 22 A shows two closely interacting protein domains that can be interpreted as the N- and C-terminal halves of the protein. The projection structure of Pgp is consistent with the recently published x-ray structure of MsbA, a lipid A flippase from Escherichia coli with high sequence homology to Pgp but only when the two MsbA subunits are rotated to bring their nucleotide binding domains together.