人源 IRSp53 与肠出血性大肠杆菌转位紧密素受体 Tir 形成复合物的结构基础。
Structural basis for complex formation between human IRSp53 and the translocated intimin receptor Tir of enterohemorrhagic E. coli.
机构信息
Division of Structural Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany.
出版信息
Structure. 2011 Sep 7;19(9):1294-306. doi: 10.1016/j.str.2011.06.015.
Abstract
Actin assembly beneath enterohemorrhagic E. coli (EHEC) attached to its host cell is triggered by the intracellular interaction of its translocated effector proteins Tir and EspF(U) with human IRSp53 family proteins and N-WASP. Here, we report the structure of the N-terminal I-BAR domain of IRSp53 in complex with a Tir-derived peptide, in which the homodimeric I-BAR domain binds two Tir molecules aligned in parallel. This arrangement provides a protein scaffold linking the bacterium to the host cell's actin polymerization machinery. The structure uncovers a specific peptide-binding site on the I-BAR surface, conserved between IRSp53 and IRTKS. The Tir Asn-Pro-Tyr (NPY) motif, essential for pedestal formation, is specifically recognized by this binding site. The site was confirmed by mutagenesis and in vivo-binding assays. It is possible that IRSp53 utilizes the NPY-binding site for additional interactions with as yet unknown partners within the host cell.
摘要
肠出血性大肠杆菌（EHEC）黏附于宿主细胞后，其转位效应蛋白 Tir 和 EspF(U)与人类 IRSp53 家族蛋白和 N-WASP 在细胞内相互作用，引发肌动蛋白聚合。本研究报告了 IRSp53 的 N 端 I-BAR 结构域与 Tir 衍生肽的复合物结构，其中同源二聚体 I-BAR 结构域结合两个平行排列的 Tir 分子。这种排列方式为细菌与宿主细胞肌动蛋白聚合机制提供了一个蛋白支架。该结构揭示了 I-BAR 表面的一个特定肽结合位点，该位点在 IRSp53 和 IRTKS 之间保守。 pedestal 形成所必需的 Tir 天冬酰胺-脯氨酸-酪氨酸（NPY）基序特异性地被该结合位点识别。该位点通过突变和体内结合测定得到证实。IRSp53 可能利用 NPY 结合位点与宿主细胞内尚未知的其他伴侣进行额外的相互作用。
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