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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Transcriptome-based systematic identification of extracellular matrix proteins
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Transcriptome-based systematic identification of extracellular matrix proteins

机译：基于转录组的细胞外基质蛋白的系统鉴定

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Extracellular matrix (ECM), which provides critical scaffolds for all adhesive cells, regulates proliferation, differentiation, and apopto-sis. Different cell types employ customized ECMs, which are thought to play important roles in the generation of so-called niches that contribute to cell-specific functions. The molecular entities of these customized ECMs, however, have not been elucidated. Here, we describe a strategy for transcriptome-wide identification of ECM proteins based on computational screening of > 60,000 full-length mouse cDNAs for secreted proteins, followed by in vitro functional assays. These assays screened the candidate proteins for ECM-assembling activities, interactions with other ECM molecules, modifications with glycosaminoglycans, and cell-adhesive activities, and were then complemented with immu-nohistochemical analysis. We identified 16 ECM proteins, of which seven were localized in basement membrane (BM) zones. The identification of these previously unknown BM proteins allowed us to construct a body map of BM proteins, which represents the comprehensive immunohistochemistry-based expression profiles of the tissue-specific customization of BMs.
机译：细胞外基质（ECM）为所有粘附细胞提供关键的支架，调节增殖，分化和凋亡。不同的细胞类型采用定制的ECM，它们被认为在有助于特定细胞功能的所谓“壁ni”的产生中发挥重要作用。但是，尚未阐明这些定制的ECM的分子实体。在这里，我们描述了基于> 60,000全长小鼠cDNAs分泌蛋白的计算筛选，然后进行体外功能测定的ECM蛋白转录组范围识别的策略。这些测定法筛选了候选蛋白质的ECM组装活性，与其他ECM分子的相互作用，糖胺聚糖修饰和细胞粘附活性，然后进行了免疫组织化学分析。我们鉴定出16种ECM蛋白，其中7种位于基底膜（BM）区。这些以前未知的BM蛋白的鉴定使我们能够构建BM蛋白的人体图，它代表了BM组织特异性定制的基于免疫组化的全面表达谱。

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