Cell surface labeling and mass spectrometry reveal diversity of cell surface markers and signaling molecules expressed in undifferentiated mouse embryonic stem cells
Cell surface labeling and mass spectrometry reveal diversity of cell surface markers and signaling molecules expressed in undifferentiated mouse embryonic stem cells
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DOI：
10.1074/mcp.m500216-mcp200
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发表时间：
2005-12-01
影响因子：
7
通讯作者：
Isobe, T
中科院分区：
文献类型：
--
作者：
Nunomura, K;Nagano, K;Isobe, T
虽然细胞表面蛋白和细胞外配体之间的相互作用是启动胚胎干细胞分化为特定细胞谱系的关键，但这些细胞的质膜蛋白组分在很大程度上是未知的。我们在这里描述了一组蛋白质的未分化的小鼠胚胎干细胞系D3的表面上表达。这些蛋白质的鉴定使用细胞表面标记与生物素，质膜的亚细胞分级分离，和基于质谱的蛋白质鉴定技术的组合。从965个独特的肽携带生物素标记，我们分配324个蛋白质，包括235个蛋白质，具有推定的信号序列和/或跨膜片段。受体、转运蛋白和细胞粘附分子是鉴定的主要蛋白质类。除了已知的胚胎干细胞的细胞表面标志物，如碱性磷酸酶，该分析还确定了59个分化相关分子簇和80多个细胞信号通路的组成部分，这些分子簇和组成部分是许多不同细胞谱系的特征。我们鉴定了白血病抑制因子、白细胞介素6和骨形态发生蛋白的受体，这些受体在维持未分化的小鼠胚胎干细胞中起关键作用。我们还鉴定了生长因子/细胞因子的受体，如成纤维细胞生长因子、血小板衍生生长因子、肝配蛋白、Hedgehog和Wnt，其为细胞分化和胚胎发育提供信号。最后，我们确定了各种整合素，细胞粘附分子和基质金属蛋白酶。这些结果表明，D3细胞表达不同的细胞表面蛋白，其功能是维持多能性，使细胞能够响应各种外部信号，启动分化为各种细胞类型。
Although interactions between cell surface proteins and extracellular ligands are key to initiating embryonic stem cell differentiation to specific cell lineages, the plasma membrane protein components of these cells are largely unknown. We describe here a group of proteins expressed on the surface of the undifferentiated mouse embryonic stem cell line D3. These proteins were identified using a combination of cell surface labeling with biotin, subcellular fractionation of plasma membranes, and mass spectrometry-based protein identification technology. From 965 unique peptides carrying biotin labels, we assigned 324 proteins including 235 proteins that have putative signal sequences and/or transmembrane segments. Receptors, transporters, and cell adhesion molecules were the major classes of proteins identified. Besides known cell surface markers of embryonic stem cells, such as alkaline phosphatase, the analysis identified 59 clusters of differentiation-related molecules and more than 80 components of multiple cell signaling pathways that are characteristic of a number of different cell lineages. We identified receptors for leukemia-inhibitory factor, interleukin 6, and bone morphogenetic protein, which play critical roles in the maintenance of undifferentiated mouse embryonic stem cells. We also identified receptors for growth factors/cytokines, such as fibroblast growth factor, platelet-derived growth factor, ephrin, Hedgehog, and Wnt, which transduce signals for cell differentiation and embryonic development. Finally we identified a variety of integrins, cell adhesion molecules, and matrix metalloproteases. These results suggest that D3 cells express diverse cell surface proteins that function to maintain pluripotency, enabling cells to respond to various external signals that initiate differentiation into a variety of cell types.