Quantitative Proteomic Analysis of Membrane Proteins Involved in Astroglial Differentiation of Neural Stem Cells by SILAC Labeling Coupled with LC-MS/MS
Quantitative Proteomic Analysis of Membrane Proteins Involved in Astroglial Differentiation of Neural Stem Cells by SILAC Labeling Coupled with LC-MS/MS
复制标题
通过 SILAC 标记结合 LC-MS/MS 对参与神经干细胞星形胶质细胞分化的膜蛋白进行定量蛋白质组分析
DOI：
10.1021/pr200677z
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发表时间：
2012-02-01
影响因子：
4.4
通讯作者：
Liang, Songping
中科院分区：
文献类型：
--
作者：
Cao, Rui;Chen, Ke;Liang, Songping
膜蛋白在神经干细胞的自我更新和分化过程中起着关键作用。在这里，我们应用SILAC（稳定同位素标记的氨基酸在细胞培养）的方法来定量比较自我更新和星形胶质细胞分化细胞的膜蛋白质组。在亚ppm质量精度的线性离子阱轨道阱仪器（LTQ-Orbitrap）的高分辨率分析导致在星形胶质细胞分化过程中超过700种不同的膜蛋白的可靠鉴定和定量。在735种定量蛋白质中，7种细胞表面蛋白质在未分化状态膜中的表达水平显著高于星形胶质细胞分化膜。转铁蛋白受体蛋白1可能成为神经干细胞表面标志物的一个新的候选者。通过对差异表达蛋白的功能聚类分析发现，在星形胶质细胞分化的神经干细胞中，大多数过表达的膜蛋白参与细胞生长、神经系统发育和能量代谢途径。总之，这项研究增加了我们对调节神经干细胞增殖和分化的复杂生物学过程的潜在机制的理解。
Membrane proteins play a critical role in the process of neural stem cell self-renewal and differentiation. Here, we apply the SILAC (stable isotope labeling by amino acids in cell culture) approach to quantitatively compare the membrane proteome of the self-renewing and the astroglial differentiating cells. High-resolution analysis on a linear ion trap-Orbitrap instrument (LTQ-Orbitrap) at sub-ppm mass accuracy resulted in confident identification and quantitation of more than 700 distinct membrane proteins during the astroglial differentiation. Of the 735 quantified proteins, seven cell surface proteins display significantly higher expression levels in the undifferentiated state membrane compared to astroglial differentiating membrane. One cell surface protein transferrin receptor protein 1 may serve as a new candidate for NSCs surface markers. Functional clustering of differentially expressed proteins by Ingenuity Pathway Analysis revealed that most of overexpressed membrane proteins in the astroglial differentiation neural stem cells are involved in cellular growth, nervous system development, and energy metabolic pathway. Taken together, this study increases our understanding of the underlying mechanisms that modulate complex biological processes of neural stem cell proliferation and differentiation.