Molecular and structural transmembrane determinants critical for embedding claudin-5 into tight junctions reveal a distinct four-helix bundle arrangement.
Molecular and structural transmembrane determinants critical for embedding claudin-5 into tight junctions reveal a distinct four-helix bundle arrangement.
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对于将claudin-5嵌入紧密连接至关重要的分子和结构跨膜决定因素揭示了独特的四螺旋束排列
DOI：
10.1042/bj20140431
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Piontek J
中科院分区：
文献类型：
--
作者：
Rossa J;Protze J;Kern C;Piontek A;Günzel D;Krause G;Piontek J
紧密连接（TJ）的组装机制和TJ链形成Cldns（claudins）的结构尚不清楚。为了确定血脑屏障相关的Cldn 3和Cldn 5组装的决定因素，通过TJ链的细胞重建和活细胞成像分析嵌合突变体。在拯救链形成缺陷突变体的基础上，我们鉴定了Cldn 5参与折叠和组装的残基（Cys 128，Ala 132，Ile 142，Ala 163，Ile 166和Leu 174）。实验结果与结构生物信息学方法相结合。最初，将实验验证的Cldn 5的ECL 2（细胞外环2）的先前模型扩展到侧翼跨膜区段（TM 3/TM 4）。一个卷曲螺旋接口可能造成交替的小和大的残基支持伴随旋钮到孔的相互作用，包括Cldn 5特定的残基在本文中确定。为了解决四螺旋束中的TM的排列，来自进化序列偶联的数据和Cldns跨膜区中分子内界面的比较建模导致了完整的Cldn 5模型。我们建议的Cldn亚型特异性分子内界面，由保守的卷曲螺旋基序和非保守残基在不同的TM位置形成的，证实了最近发布的晶体结构的Cldn 15。所鉴定的分子和结构决定因素基本上有助于将Cldns组装成TJ链。
The mechanism of TJ (tight junction) assembly and the structure of TJ strand-forming Cldns (claudins) are unclear. To identify determinants of assembly of blood–brain barrier-related Cldn3 and Cldn5, chimaeric mutants were analysed by cellular reconstitution of TJ strands and live-cell imaging. On the basis of the rescue of mutants deficient for strand formation, we identified Cldn5 residues (Cys128, Ala132, Ile142, Ala163, Ile166and Leu174) involved in Cldn folding and assembly. Experimental results were combined with structural bioinformatics approaches. Initially the experimentally validated previous model of the ECL2 (extracellular loop 2) of Cldn5 was extended to the flanking transmembrane segments (TM3/TM4). A coiled-coil interface probably caused by alternating small and large residues is supported by concomitant knob-into-hole interactions including Cldn5-specific residues identified in the present paper. To address arrangement of the TMs in a four-helix bundle, data from evolutionary sequence couplings and comparative modelling of intramolecular interfaces in the transmembrane region of Cldns led to a complete Cldn5 model. Our suggested Cldn subtype-specific intramolecular interfaces that are formed by conserved coiled-coil motifs and non-conserved residues in distinct TM positions were confirmed by the recently released crystal structure of Cldn15. The identified molecular and structural determinants essentially contribute to assembly of Cldns into TJ strands.
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影响因子：
5.5
作者：
Colegio, OR;Van Itallie, C;Anderson, JM
通讯作者：
Anderson, JM
影响因子：
3.4
作者：
Milatz, Susanne;Krug, Susanne M.;Fromm, Michael
通讯作者：
Fromm, Michael
影响因子：
15.9
作者：
Kausalya, PJ;Amasheh, S;Hunziker, W
通讯作者：
Hunziker, W
Wnt/β-catenin信号传导控制血脑屏障的发展。
影响因子：
7.8
作者：
Liebner, Stefan;Corada, Monica;Bangsow, Thorsten;Babbage, Jane;Taddei, Andrea;Czupalla, Cathrin J.;Reis, Marco;Felici, Angelina;Wolburg, Hartwig;Fruttiger, Marcus;Taketo, Makoto M.;von Melchner, Harald;Plate, Karl Heinz;Gerhardt, Holger;Dejana, Elisabetta
通讯作者：
Dejana, Elisabetta
DOI：
10.1093/protein/12.11.953
发表时间：
1999-11-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Mitaku, S;Hirokawa, T
通讯作者：
Hirokawa, T