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Preprint
Proximity Labeling Expansion Microscopy (PL-ExM) resolves structure of the interactome
bioRxiv
13 Nov 2023
PMID: 38014020
Abstract
Elucidating the spatial relationships within the protein interactome is pivotal to understanding the organization and regulation of protein-protein interactions. However, capturing the 3D architecture of the interactome presents a dual challenge: precise interactome labeling and super-resolution imaging. To bridge this gap, we present the Proximity Labeling Expansion Microscopy (PL-ExM). This innovation combines proximity labeling (PL) to spatially biotinylate interacting proteins with expansion microscopy (ExM) to increase imaging resolution by physically enlarging cells. PL-ExM unveils intricate details of the 3D interactome’s spatial layout in cells using standard microscopes, including confocal and Airyscan. Multiplexing PL-ExM imaging was achieved by pairing the PL with immunofluorescence staining. These multicolor images directly visualize how interactome structures position specific proteins in the protein-protein interaction network. Furthermore, PL-ExM stands out as an assessment method to gauge the labeling radius and efficiency of different PL techniques. The accuracy of PL-ExM is validated by our proteomic results from PL mass spectrometry. Thus, PL-ExM is an accessible solution for 3D mapping of the interactome structure and an accurate tool to access PL quality.
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Details
- Title
- Proximity Labeling Expansion Microscopy (PL-ExM) resolves structure of the interactome
- Creators
- Sohyeon Park - University of California, IrvineXiaorong Wang - University of California, IrvineXiangpeng Li - University of California, San FranciscoXiao Huang - Drexel UniversityKatie C. Fong - California State University Los AngelesClinton Yu - University of California, IrvineArthur A. Tran - University of California, San FranciscoLorenzo Scipioni - Fluor (United States)Zhipeng Dai - University of California, IrvineLan Huang - University of California, IrvineXiaoyu Shi - University of California, Irvine
- Publication Details
- bioRxiv
- Publisher
- Cold Spring Harbor Laboratory
- Edition
- 1.1
- Number of pages
- 25
- Resource Type
- Preprint
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems
- Other Identifier
- 991022130762804721