Department of Medical Protein Research, VIB, Ghent, Belgium.
PLoS One. 2012;7(9):e44143. doi: 10.1371/journal.pone.0044143. Epub 2012 Sep 10.
The mammalian two-hybrid system MAPPIT allows the detection of protein-protein interactions in intact human cells. We developed a random mutagenesis screening strategy based on MAPPIT to detect mutations that disrupt the interaction of one protein with multiple protein interactors simultaneously. The strategy was used to detect residues of the human cytidine deaminase Apobec3G that are important for its homodimerization and its interaction with the HIV-1 Gag and Vif proteins. The strategy is able to identify the previously described head-to-head homodimerization interface in the N-terminal domain of Apobec3G. Our analysis further detects two new potential interaction surfaces in the N-and C-terminal domain of Apobec3G for interaction with Vif and Gag or for Apobec3G dimerization.
哺乳动物双杂交系统 MAPPIT 允许在完整的人类细胞中检测蛋白质-蛋白质相互作用。我们开发了一种基于 MAPPIT 的随机诱变筛选策略，以检测同时破坏一种蛋白质与多个蛋白质相互作用子相互作用的突变。该策略用于检测人胞苷脱氨酶 Apobec3G 的重要残基，这些残基对于其同源二聚体化及其与 HIV-1 Gag 和 Vif 蛋白的相互作用至关重要。该策略能够识别 Apobec3G N 端结构域中先前描述的头对头同源二聚化界面。我们的分析进一步检测到 Apobec3G 的 N 和 C 端结构域中的两个新的潜在相互作用表面，用于与 Vif 和 Gag 相互作用或用于 Apobec3G 二聚化。
