人类PDZ结构域蛋白组2.0：一种用于通过酵母双杂交检测PDZ相互作用的新资源的特性分析

The Human PDZome 2.0: Characterization of a New Resource to Test for PDZ Interactions by Yeast Two-Hybrid.

作者信息

Castro-Cruz Monica, Lembo Frédérique, Borg Jean-Paul, Travé Gilles, Vincentelli Renaud, Zimmermann Pascale

机构信息

Department of Human Genetics, KU Leuven, 3000 Leuven, Belgium.

Équipe Labellisée Ligue 2018, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix-Marseille Université, 13009 Marseille, France.

出版信息

Membranes (Basel). 2023 Aug 17;13(8):737. doi: 10.3390/membranes13080737.

DOI:10.3390/membranes13080737

PMID:37623798

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10456741/

Abstract

PSD95-disc large-zonula occludens (PDZ) domains are globular modules of 80-90 amino acids that co-evolved with multicellularity. They commonly bind to carboxy-terminal sequences of a plethora of membrane-associated proteins and influence their trafficking and signaling. We previously built a PDZ resource (PDZome) allowing us to unveil human PDZ interactions by Yeast two-hybrid. Yet, this resource is incomplete according to the current knowledge on the human PDZ proteome. Here we built the PDZome 2.0 library for Yeast two-hybrid, based on a PDZ library manually curated from online resources. The PDZome2.0 contains 305 individual clones (266 PDZ domains in isolation and 39 tandems), for which all boundaries were designed based on available PDZ structures. Using as bait the E6 oncoprotein from HPV16, a known promiscuous PDZ interactor, we show that PDZome 2.0 outperforms the previous resource.

摘要

PSD95-盘状大闭合小带（PDZ）结构域是由80-90个氨基酸组成的球状模块，与多细胞生物共同进化。它们通常与大量膜相关蛋白的羧基末端序列结合，并影响其运输和信号传导。我们之前构建了一个PDZ资源库（PDZome），使我们能够通过酵母双杂交揭示人类PDZ相互作用。然而，根据目前关于人类PDZ蛋白质组的知识，这个资源库并不完整。在这里，我们基于从在线资源中手动筛选的PDZ文库，构建了用于酵母双杂交的PDZome 2.0文库。PDZome2.0包含305个单独的克隆（266个孤立的PDZ结构域和39个串联体），其所有边界均根据可用的PDZ结构设计。使用来自HPV16的E6癌蛋白作为诱饵，E6是一种已知的多价PDZ相互作用蛋白，我们表明PDZome 2.0优于之前的资源。