Both the N- and C- terminal regions of the Chlamydial inclusion protein D (IncD) are required for interaction with the pleckstrin homology domain of the ceramide transport protein CERT.
Both the N- and C- terminal regions of the Chlamydial inclusion protein D (IncD) are required for interaction with the pleckstrin homology domain of the ceramide transport protein CERT.
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DOI：
10.1016/j.bbrc.2018.09.168
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发表时间：
2018-11-10
影响因子：
3.1
通讯作者：
Hanada K
中科院分区：
文献类型：
--
作者：
Kumagai K;Elwell CA;Ando S;Engel JN;Hanada K
专性细胞内细菌沙眼衣原体需要宿主脂质神经酰胺在细胞内膜腔内进行复制。衣原体包膜蛋白D （IncD）由两个紧密相连的长疏水结构域组成，其N端和c端暴露于宿主细胞质，直接与神经酰胺运输蛋白（CERT）的pleckstrin同源结构域（PH）结合，可能将神经酰胺重定向到包膜上。这种相互作用所需的IncD的精确区域尚未划定。通过共转染研究和系统发育研究，我们证明了IncD N端和c端区域都是与CERT PH结构域结合并定义关键相互作用残基所必需的。原生凝胶电泳分析表明，IncD的跨膜区域形成了抗sds但对二硫苏糖醇敏感的同二聚体，这些同二聚体又可以通过额外的N端和c端结构域接触组装成高阶低聚物。我们认为IncD寡聚化可能促进了与CERT的高亲和力结合，允许沙眼衣原体有效地将宿主神经酰胺重定向到包涵体上。
An obligate intracellular bacterium Chlamydia trachomatis requires host lipid ceramide for their replication within an intracellular membranous compartment, the inclusion. Chlamydial inclusion membrane protein D (IncD) composed of two closely linked long hydrophobic domains with their N- and C-termini exposed to the host cytosol, binds directly to the pleckstrin homology (PH) domain of the ceramide transport protein (CERT), likely redirecting ceramide to the inclusion. The precise regions of IncD required for this interaction have not been delineated. Using co-transfection studies together with phylogenetic studies, we demonstrate that both the IncD N- and C-terminal regions are required for binding to the CERT PH domain and define key interaction residues. Native gel electrophoresis analysis demonstrates that the transmembrane region of IncD forms SDS-resistant but dithiothreitol-sensitive homodimers, which in turn can assemble to form higher order oligomers through additional N- and C-terminal domain contacts. We propose that IncD oligomerization may facilitate high affinity binding to CERT, allowing C. trachomatis to efficiently redirect host ceramide to the inclusion.
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