Key Laboratory of Protein Chemistry and Developmental Biology of Education Committee, College of Life Sciences, Hunan Normal University , Changsha 410081, PR China.
J Proteome Res. 2012 Feb 3;11(2):829-38. doi: 10.1021/pr200677z. Epub 2012 Jan 5.
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.
膜蛋白在神经干细胞自我更新和分化过程中起着至关重要的作用。在这里，我们应用 SILAC（稳定同位素标记的氨基酸细胞培养）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-Orbitrap 仪器（LTQ-Orbitrap）上进行高分辨率分析，在星形胶质细胞分化过程中鉴定和定量了超过 700 种不同的膜蛋白。在 735 种定量蛋白中，7 种细胞表面蛋白在未分化状态的膜中表达水平明显高于星形胶质细胞分化的膜。一种细胞表面蛋白转铁蛋白受体蛋白 1 可能作为神经干细胞表面标志物的新候选物。通过 IPA 进行的差异表达蛋白功能聚类分析表明，在星形胶质细胞分化的神经干细胞中过度表达的大多数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复杂生物学过程的潜在机制的理解。
