Mechanism of intermediate filament recognition by plakin repeat domains revealed by envoplakin targeting of vimentin.
Mechanism of intermediate filament recognition by plakin repeat domains revealed by envoplakin targeting of vimentin.
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DOI：
10.1038/ncomms10827
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发表时间：
2016-03-03
影响因子：
16.6
通讯作者：
Chidgey M
中科院分区：
文献类型：
--
作者：
Fogl C;Mohammed F;Al-Jassar C;Jeeves M;Knowles TJ;Rodriguez-Zamora P;White SA;Odintsova E;Overduin M;Chidgey M
斑块蛋白形成细胞连接和细胞骨架之间的关键连接;它们在上皮细胞和心肌细胞内的破坏导致皮肤起泡疾病和心肌病。Envoplakin有一个单一的plakin重复结构域（PRD），它通过一个未解决的机制识别中间丝。在这里，我们报告的晶体结构的envoplakin的完整的PRD折叠，揭示其正电结合沟内的结合决定簇。它的五个内部重复序列中的四个通过核磁共振光谱鉴定的五个基本决定簇识别波形蛋白内的带负电荷的斑块。Lys 1901或Arg 1914结合决定簇的突变使培养细胞中的细胞内波形蛋白和角蛋白丝的异二聚体envoplakin离域。当其残基Asp 112或Asp 119突变时，波形蛋白的识别被消除。后者槽中间丝杆到基本PRD域槽通过电空间互补性在一个广泛适用的机制。这揭示了plakin家族成员如何与细胞骨架框架形成动态联系。 斑块蛋白将细胞连接到细胞骨架框架，并且它们在上皮细胞和心肌细胞内的破坏引起皮肤起泡疾病和心肌病。在这里，作者使用结构生物学的方法来揭示机制，允许斑蛋白识别细胞骨架内的中间丝。
Plakin proteins form critical connections between cell junctions and the cytoskeleton; their disruption within epithelial and cardiac muscle cells cause skin-blistering diseases and cardiomyopathies. Envoplakin has a single plakin repeat domain (PRD) which recognizes intermediate filaments through an unresolved mechanism. Herein we report the crystal structure of envoplakin's complete PRD fold, revealing binding determinants within its electropositive binding groove. Four of its five internal repeats recognize negatively charged patches within vimentin via five basic determinants that are identified by nuclear magnetic resonance spectroscopy. Mutations of the Lys1901 or Arg1914 binding determinants delocalize heterodimeric envoplakin from intracellular vimentin and keratin filaments in cultured cells. Recognition of vimentin is abolished when its residues Asp112 or Asp119 are mutated. The latter slot intermediate filament rods into basic PRD domain grooves through electrosteric complementarity in a widely applicable mechanism. Together this reveals how plakin family members form dynamic linkages with cytoskeletal frameworks. Plakin proteins link cell junctions to cytoskeletal frameworks, and their disruption within epithelial and cardiac muscle cells cause skin blistering diseases and cardiomyopathies. Here the authors use structural biology approaches to reveal the mechanism that allows plakins to recognize intermediate filaments within the cytoskeleton.