Qianqian Jiang, Bo Jiang, Yukui Zhang, Lihua Zhang
State Key Laboratory of Medical Proteomics, National Chromatographic R. & A. Center, CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
University of Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2025;2908:111-123. doi: 10.1007/978-1-0716-4434-8_8.
As a central platform for exchange of signaling communication and substance between cells and within cells, mapping of membrane proteome can provide new insights into the pathophysiology of cancer and other diseases, explore potential drug targets and biomarkers for prognosis or diagnosis, and ultimately contribute to the development of precision medicine. However, mass spectrometry-based identification of membrane proteins poses many challenges due to the intrinsic properties of membrane proteins, especially high hydrophobicity and low abundance. Herein, we described a metal organic layers (MOL)-based sample preparation method for membrane proteome analysis in living cells, involving the cell surface engineering process through the exquisite interaction between MOL and phospholipid bilayers on the membrane and the following biomembrane fusion process, providing a simple and robust tool for dissection of membrane proteome with high specificity. This chapter details the preparation of such metal organic layers and enrichment workflow of HeLa membrane proteins.
作为细胞间以及细胞内信号通信和物质交换的核心平台，膜蛋白质组图谱能够为癌症及其他疾病的病理生理学提供新见解，探索潜在的药物靶点以及用于预后或诊断的生物标志物，并最终推动精准医学的发展。然而，由于膜蛋白的固有特性，尤其是高疏水性和低丰度，基于质谱的膜蛋白鉴定面临诸多挑战。在此，我们描述了一种基于金属有机层（MOL）的活细胞中膜蛋白质组分析的样品制备方法，该方法涉及通过MOL与膜上磷脂双层之间的精细相互作用进行细胞表面工程处理以及随后的生物膜融合过程，为高特异性剖析膜蛋白质组提供了一种简单且可靠的工具。本章详细介绍了此类金属有机层的制备以及HeLa膜蛋白的富集工作流程。
