Interaction of the profilaggrin N-terminal domain with loricrin in human cultured keratinocytes and epidermis.
Interaction of the profilaggrin N-terminal domain with loricrin in human cultured keratinocytes and epidermis.
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DOI：
10.1038/jid.2011.460
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发表时间：
2012-04
期刊：
影响因子：
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通讯作者：
K. Yoneda;T. Nakagawa;O. Lawrence;J. Huard;T. Demitsu;Y. Kubota;R. Presland
中科院分区：
文献类型：
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作者：
K. Yoneda;T. Nakagawa;O. Lawrence;J. Huard;T. Demitsu;Y. Kubota;R. Presland
两个共表达的分化标志物，原丝聚蛋白和兜甲蛋白之间的关系目前尚不清楚。在本研究中，我们探讨了在角质形成细胞和表皮中的微丝聚集蛋白原N-末端结构域（PND）与兜甲蛋白的相互作用。人表皮共聚焦免疫荧光显微镜分析显示PND与兜甲蛋白共定位。兜甲蛋白被核转染到HaCaT细胞中，与PND共定位于细胞核和细胞质中。PND定位于表皮颗粒层细胞的细胞核和细胞质。核转染的PND还与角蛋白10（K10）共定位于细胞核和细胞质中。人表皮的免疫电镜分析证实了核转染角质形成细胞中的发现。酵母双杂交实验表明，人和小鼠PND的B结构域与兜甲蛋白相互作用。谷胱甘肽S-转移酶（GST）下拉分析使用重组GST-PND显示PND与兜甲蛋白和K10相互作用。在器官型皮肤培养模型中PND的敲除导致聚丝蛋白表达的损失和透明角质蛋白颗粒的大小和数量的减少，以及兜甲蛋白的表达显著减少。考虑到PND的表达与角质形成细胞终末分化密切相关，我们得出结论，PND与兜甲蛋白和K10在体内相互作用，这些相互作用可能与皮质包膜组装和随后的表皮屏障形成有关。
The relationship between the two coexpressed differentiation markers, profilaggrin and loricrin, is not clear right now. In this study, we explored the interaction of profilaggrinN-terminal domain (PND) with loricrin in keratinocytes and epidermis. Confocal immunofluorescence microscopic analysis of human epidermis showed that PND colocalized with loricrin. Loricrin nucleofected into HaCaT cells colocalized with PND in the nucleus and cytoplasm. The PND localizes to both the nucleus and cytoplasm of epidermal granular layer cells. Nucleofected PND also colocalized with keratin 10 (K10) in the nucleus and cytoplasm. Immunoelectron microscopic analysis of human epidermis confirmed the findings in nucleofected keratinocytes. Yeast two-hybrid assays showed that the B domain of human and mouse PND interacted with loricrin. The glutathioneS-transferase (GST) pull-down analysis using recombinant GST-PND revealed that PND interacted with loricrin and K10. Knockdown of PND in an organotypic skin culture model caused loss of filaggrin expression and a reduction in both the size and number of keratohyalin granules, as well as markedly reduced expression of loricrin. Considering that expression of PND is closely linked to keratinocyte terminal differentiation, we conclude that PND interacts with loricrin and K10in vivoand that these interactions are likely to be relevant for cornified envelope assembly and subsequent epidermal barrier formation.