Issue 34, 2024

Proximity labeling expansion microscopy (PL-ExM) evaluates interactome labeling techniques

Abstract

Understanding protein–protein interactions (PPIs) through proximity labeling has revolutionized our comprehension of cellular mechanisms and pathology. Various proximity labeling techniques, such as HRP, APEX, BioID, TurboID, and μMap, have been widely used to biotinylate PPIs or organelles for proteomic profiling. However, the variability in labeling precision and efficiency of these techniques often results in limited reproducibility in proteomic detection. We address this persistent challenge by introducing proximity labeling expansion microscopy (PL-ExM), a super-resolution imaging technique that combines expansion microscopy with proximity labeling techniques. PL-ExM enabled up to 17 nm resolution with microscopes widely available, providing visual comparison of the labeling precision, efficiency, and false positives of different proximity labeling methods. Our mass spectrometry proteomic results confirmed that PL-ExM imaging is reliable in guiding the selection of proximity labeling techniques and interpreting the proteomic results with new spatial information.

Graphical abstract: Proximity labeling expansion microscopy (PL-ExM) evaluates interactome labeling techniques

Supplementary files

Article information

Article type
Paper
Submitted
11 mar. 2024
Accepted
22 jul. 2024
First published
30 jul. 2024

J. Mater. Chem. B, 2024,12, 8335-8348

Proximity labeling expansion microscopy (PL-ExM) evaluates interactome labeling techniques

S. Park, X. Wang, Y. Mo, S. Zhang, X. Li, K. C. Fong, C. Yu, A. A. Tran, L. Scipioni, Z. Dai, X. Huang, L. Huang and X. Shi, J. Mater. Chem. B, 2024, 12, 8335 DOI: 10.1039/D4TB00516C

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