A modification of the split-tobacco etch virus method for monitoring interactions between membrane proteins in mammalian cells

被引：6

作者：

Capdevila-Nortes, Xavier ^{[1
]}

Lopez-Hernandez, Tania ^{[1
]}

Ciruela, Francisco ^{[2
]}

Estevez, Raul ^{[1
,3
]}

机构：

[1] Univ Barcelona, Fac Med, Dept Ciencias Fisiol 2, Secc Fisiol, Barcelona 08907, Spain

[2] Univ Barcelona, Fac Med, Dept Patol & Terapeut Expt, Unitat Farmacol, Barcelona 08907, Spain

[3] Ctr Invest Red Enfermedades Rara CIBERER, U750, Barcelona 08907, Spain

来源：

ANALYTICAL BIOCHEMISTRY | 2012年 / 423卷 / 01期

关键词：

Membrane proteins; Protein interaction; Split-TEV; MEGALENCEPHALIC LEUKOENCEPHALOPATHY; DESIGN; ASSAY; MLC1;

D O I：

10.1016/j.ab.2012.01.022

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Despite progress in the development of methods to monitor protein interactions, studies of interactions between membrane proteins in mammalian cells remain challenging. Protein complementation assays (PCAs) are commonly used to study interactions between proteins due to their simplicity. They are based on interaction-mediated reconstitution of a reporter protein, which can be easily monitored. Recently, a protein complementation method named split-TEV (tobacco etch virus) has been developed and is based on the functional reconstitution of TEV protease and subsequent proteolytic-mediated activation of reporters. In this work, we have developed a modification of the split-TEV method to study the interactions between membrane proteins with increased specificity. This assay was validated by addressing the interactions between different membrane proteins, including G protein-coupled receptors (GPCRs) and ion channels. By comparing it with another PCA, we found that this new method showed a higher sensitivity. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：109 / 118

页数：10

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