Purification of eukaryotic tetherin/Vpu proteins and detection of their interaction by ELISA
Purification of eukaryotic tetherin/Vpu proteins and detection of their interaction by ELISA
复制标题
真核系链蛋白/Vpu 蛋白的纯化并通过 ELISA 检测其相互作用
DOI：
10.1016/j.pep.2013.07.015
复制
发表时间：
2013-10-01
影响因子：
1.6
通讯作者：
Yu, Xianghui
中科院分区：
文献类型：
--
作者：
Lv, Mingyu;Zhu, Yingzi;Yu, Xianghui
Tetherin/BST-2/CD 317抑制HIV-1从感染细胞释放，而HIV-1 Vpu基于每种蛋白质的跨膜结构域之间的分子间相互作用有效地拮抗Tetherin。在这项研究中，我们成功地部分纯化了His标记的tetherin与glycophosphatidylinositol删除（delGPI）和His标记的全长Vpu从瞬时转染的293 T细胞使用亲和层析。通过pull-down试验和ELISA观察这些纯化蛋白之间的体外相互作用。通过ELISA检测Vpu/tetherin相互作用是一种新的方法，将有利于体外抑制剂筛选。成功的共纯化的tetherin/Vpu复合物也提供了进一步的结构研究的基础。(C)2013 Elsevier Inc. All rights reserved.
Tetherin/BST-2/CD317 inhibits HIV-1 release from infected cells, while HIV-1 Vpu efficiently antagonizes tetherin based on intermolecular interactions between the transmembrane domains of each protein. In this study, we successfully partially purified His-tagged tetherin with a glycophosphatidylinositol deletion (delGPI) and His-tagged full-length Vpu from transiently transfected 293T cells using affinity chromatography. The in vitro interaction between these purified proteins was observed by a pull-down assay and ELISA. Detection of the Vpu/tetherin interaction by ELISA is a novel approach that would be advantageous for inhibitor screening in vitro. Successful co-purification of the tetherin/Vpu complex also provides a basis for further structural studies. (C) 2013 Elsevier Inc. All rights reserved.