Quantitative proteome analysis of alveolar type-II cells reveals a connection of integrin receptor subunits beta 2/6 and WNT signaling.

Quantitative proteome analysis of alveolar type-II cells reveals a connection of integrin receptor subunits beta 2/6 and WNT signaling.
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II 型肺泡细胞的定量蛋白质组分析揭示了整合素受体亚基 β 2/6 与 WNT 信号传导之间的联系

DOI:
10.1021/pr400573k
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发表时间:
2013
影响因子:
4.4
通讯作者:
Preissner
Preissner
中科院分区:
生物学2区
文献类型:
--
作者:
Mukhametshina;Contreras;Ahlbrecht;Carraro;Cabrera-Fuentes;Voswinckel;Seeger;Bellusci;Scharffetter-Kochanek;Bagaeva;Preissner

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肺泡II型细胞(ATII细胞)是肺祖细胞,负责肺泡上皮细胞的再生过程中的稳态转换和响应损伤。ATII细胞的表征将对我们理解和治疗肺部疾病产生深远的影响。新的ATII细胞表面蛋白的鉴定可用于这些细胞的分选和富集以用于进一步表征。在这里,我们结合了一个高分辨率的质谱为基础的膜蛋白质组学方法,使用肺的SILAC小鼠与一个基于微阵列的ATII细胞的转录组分析。我们鉴定了16种蛋白质,这些蛋白质在ATII细胞的膜部分中富集,并且其基因在这些细胞中高度表达。有趣的是,我们通过qRT-PCR表达分析和蛋白质提取物的Western印迹分析证实了其中两个基因整合素β 2和6(Itgb 2和Itgb 6)的数据。此外,流式细胞术和免疫组织化学在成人肺显示,ITGB 2和ITGB 6存在于亚群的表面活性剂相关蛋白C阳性细胞,表明存在不同类型的ATII细胞。此外,对Itgb 2-/-小鼠的分析表明,Itgb 2是肺中适当WNT信号调节所必需的。
Alveolar type-II cells (ATII cells) are lung progenitor cells responsible for regeneration of alveolar epithelium during homeostatic turnover and in response to injury. Characterization of ATII cells will have a profound impact on our understanding and treatment of lung disease. The identification of novel ATII cell-surface proteins can be used for sorting and enrichment of these cells for further characterization. Here we combined a high-resolution mass spectrometry-based membrane proteomic approach using lungs of the SILAC mice with an Affymetrix microarray-based transcriptome analysis of ATII cells. We identified 16 proteins that are enriched in the membrane fraction of ATII cells and whose genes are highly expressed in these cells. Interestingly, we confirmed our data for two of these genes, integrin beta 2 and 6 (Itgb2andItgb6), by qRT-PCR expression analysis and Western blot analysis of protein extracts. Moreover, flow cytometry and immunohistochemistry in adult lung revealed that ITGB2 and ITGB6 are present in subpopulations of surfactant-associated-protein-C-positive cells, suggesting the existence of different types of ATII cells. Furthermore, analysis of theItgb2–/–mice showed thatItgb2is required for proper WNT signaling regulation in the lung.
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