Issue 42, 2023

Metal organic layers enabled cell surface engineering coupling biomembrane fusion for dynamic membrane proteome profiling

Abstract

Systematically dissecting the highly dynamic and tightly communicating membrane proteome of living cells is essential for the system-level understanding of fundamental cellular processes and intricate relationship between membrane-bound organelles constructed through membrane traffic. While extensive efforts have been made to enrich membrane proteins, their comprehensive analysis with high selectivity and deep coverage remains a challenge, especially at the living cell state. To address this problem, we developed the cell surface engineering coupling biomembrane fusion method to map the whole membrane proteome from the plasma membrane to various organelle membranes taking advantage of the exquisite interaction between two-dimensional metal–organic layers and phospholipid bilayers on the membrane. This approach, which bypassed conventional biochemical fractionation and ultracentrifugation, facilitated the enrichment of membrane proteins in their native phospholipid bilayer environment, helping to map the membrane proteome with a specificity of 77% and realizing the deep coverage of the HeLa membrane proteome (5087 membrane proteins). Furthermore, membrane N-phosphoproteome was profiled by integrating the N-phosphoproteome analysis strategy, and the dynamic membrane proteome during apoptosis was deciphered in combination with quantitative proteomics. The features of membrane protein N-phosphorylation modifications and many differential proteins during apoptosis associated with mitochondrial dynamics and ER homeostasis were found. The method provided a simple and robust strategy for efficient analysis of membrane proteome, offered a reliable platform for research on membrane-related cell dynamic events and expanded the application of metal–organic layers.

Graphical abstract: Metal organic layers enabled cell surface engineering coupling biomembrane fusion for dynamic membrane proteome profiling

Supplementary files

Article information

Article type
Edge Article
Submitted
19 7 2023
Accepted
30 9 2023
First published
05 10 2023
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2023,14, 11727-11736

Metal organic layers enabled cell surface engineering coupling biomembrane fusion for dynamic membrane proteome profiling

Q. Jiang, H. Wang, Z. Qiao, Y. Hou, Z. Sui, B. Zhao, Z. Liang, B. Jiang, Y. Zhang and L. Zhang, Chem. Sci., 2023, 14, 11727 DOI: 10.1039/D3SC03725H

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