Instituto de Biología Molecular y Celular del Cáncer, Consejo Superior de Investigaciones Científicas - University of Salamanca, Campus Unamuno, 37007 Salamanca, Spain.
Centro Universitario de la Defensa, Ctra. Huesca s/n, 50090 Zaragoza, Spain.
Structure. 2019 Jun 4;27(6):952-964.e6. doi: 10.1016/j.str.2019.03.016. Epub 2019 Apr 18.
Mechanical stability of epithelia requires firm attachment to the basement membrane via hemidesmosomes. Dysfunction of hemidesmosomal proteins causes severe skin-blistering diseases. Two plakins, plectin and BP230 (BPAG1e), link the integrin α6β4 to intermediate filaments in epidermal hemidesmosomes. Here, we show that a linear sequence within the isoform-specific N-terminal region of BP230 binds to the third and fourth FnIII domains of β4. The crystal structure of the complex and mutagenesis analysis revealed that BP230 binds between the two domains of β4. BP230 induces closing of the two FnIII domains that are locked in place by an interdomain ionic clasp required for binding. Disruption of BP230-β4 binding prevents recruitment of BP230 to hemidesmosomes in human keratinocytes, revealing a key role of this interaction for hemidesmosome assembly. Phosphomimetic substitutions in β4 and BP230 destabilize the complex. Thus, our study provides insights into the architecture of hemidesmosomes and potential mechanisms of regulation.
上皮组织的机械稳定性需要通过半桥粒牢固地附着在基底膜上。半桥粒蛋白功能障碍会导致严重的皮肤水疱病。两种斑联蛋白,即网蛋白和 BP230(BPAG1e),将整合素α6β4连接到表皮半桥粒中的中间丝上。在这里,我们表明 BP230 异构体特异性 N 端区域内的线性序列与β4 的第三和第四 FnIII 结构域结合。复合物的晶体结构和突变分析表明,BP230 结合在β4 的两个结构域之间。BP230 诱导两个 FnIII 结构域闭合,两个结构域被结合所必需的结构域内离子扣锁定。BP230-β4 结合的破坏阻止了 BP230 在人角质形成细胞中向半桥粒的募集,揭示了这种相互作用对半桥粒组装的关键作用。β4 和 BP230 中的磷酸模拟取代使复合物不稳定。因此,我们的研究提供了对半桥粒结构和潜在调节机制的深入了解。