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 systems-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, the comprehensive analysis of them 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 plasma membrane to various organelle membranes taking advantage of the exquisite interaction between two-dimensional metal-organic layers and phospholipid bilayers on membrane. This approach, which bypassed conventional biochemical fractionation and ultracentrifugation, facilitated the enrichment of membrane proteins in their native phospholipid bilayer environment, helping map the membrane proteome with a specificity of 77% and realizing the deep coverage of the HeLa membrane proteome (5,087 membrane proteins). Furthermore, membrane N-phosphoproteome was profiled by integrating N-phosphoproteome analysis strategy and 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, 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.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Velos, Orbitrap Exploris 480, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Malignant Cell

DISEASE(S): Disease Free

SUBMITTER: Qianqian Jiang  

LAB HEAD: Qianqian Jiang

PROVIDER: PXD044945 | Pride | 2024-01-26

REPOSITORIES: Pride