Boeke Dominik, Trautmann Susanne, Meurer Matthias, Wachsmuth Malte, Godlee Camilla, Knop Michael, Kaksonen Marko
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH) Deutsches Krebsforschungszentrum (DKFZ) DKFZ-ZMBH-Allianz, Heidelberg, Germany.
Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH) Deutsches Krebsforschungszentrum (DKFZ) DKFZ-ZMBH-Allianz, Heidelberg, Germany.
Mol Syst Biol. 2014 Nov 3;10(11):756. doi: 10.15252/msb.20145422.
Clathrin-mediated endocytosis is a highly conserved intracellular trafficking pathway that depends on dynamic protein-protein interactions between up to 60 different proteins. However, little is known about the spatio-temporal regulation of these interactions. Using fluorescence (cross)-correlation spectroscopy in yeast, we tested 41 previously reported interactions in vivo and found 16 to exist in the cytoplasm. These detected cytoplasmic interactions included the self-interaction of Ede1, homolog of mammalian Eps15. Ede1 is the crucial scaffold for the organization of the early stages of endocytosis. We show that oligomerization of Ede1 through its central coiled coil domain is necessary for its localization to the endocytic site and we link the oligomerization of Ede1 to its function in locally concentrating endocytic adaptors and organizing the endocytic machinery. Our study sheds light on the importance of the regulation of protein-protein interactions in the cytoplasm for the assembly of the endocytic machinery in vivo.
网格蛋白介导的内吞作用是一种高度保守的细胞内运输途径，它依赖于多达60种不同蛋白质之间动态的蛋白质-蛋白质相互作用。然而，对于这些相互作用的时空调节知之甚少。我们利用酵母中的荧光（交叉）相关光谱技术，在体内测试了41种先前报道的相互作用，发现其中16种存在于细胞质中。这些检测到的细胞质相互作用包括哺乳动物Eps15的同源物Ede1的自身相互作用。Ede1是内吞作用早期阶段组织的关键支架。我们表明，Ede1通过其中心卷曲螺旋结构域的寡聚化对于其定位于内吞位点是必要的，并且我们将Ede1的寡聚化与其在局部浓缩内吞衔接蛋白和组织内吞机制中的功能联系起来。我们的研究揭示了细胞质中蛋白质-蛋白质相互作用的调节对于体内内吞机制组装的重要性。
