A new version of this entry is available:
Loading...
Article
2025
An enzymatic cascade enables sensitive and specific proximity labeling proteomics in challenging biological systems
An enzymatic cascade enables sensitive and specific proximity labeling proteomics in challenging biological systems
Abstract (English)
Ascorbate peroxidase (APEX) is a proximity labeling enzyme used for subcellular proteomics at high spatial and temporal resolution. However, toxicity of its substrate hydrogen peroxide and background labeling by endogenous peroxidases limit its use to in vitro studies of specific cell types. To minimize toxicity and reduce non-specific background labeling we establish a more versatile in situ APEX activation (iAPEX) workflow by combining APEX2 with a D-amino acid oxidase to locally produce hydrogen peroxide. Using iAPEX, we profile the proteomes of a cellular microdomain, the primary cilium, in cell lines not readily accessible to conventional APEX labeling and identify unknown ciliary proteins. Our study validates common ciliary proteins across two distinct cell lines, while observed differences may reflect heterogeneity in primary cilia proteomes. Furthermore, iAPEX proximity labeling is applicable to a range of cellular compartments including mitochondria and lipid droplets and can be employed in Xenopus laevis , which provides a proof-of-concept for future in vivo applications.
File is subject to an embargo until
This is a correction to:
A correction to this entry is available:
This is a new version of:
Other version
Notes
Publication license
Publication series
Published in
Nature Communications, 16 (2025), 1, 9691.
https://doi.org/10.1038/s41467-025-65405-8.
ISSN: 2041-1723
London : Nature Publishing Group UK
Other version
Faculty
Institute
Examination date
Supervisor
Cite this publication
Sroka, T. J., Sanwald, L. K., Prasai, A., Hoeren, J., Trivigno, V., Chaumet, V., Krauß, L. M., Weber, D., Yildiz, D., von der Malsburg, K., Walentek, P., Haberkant, P., Schrul, B., Feistel, K., & Mick, D. U. (2025). An enzymatic cascade enables sensitive and specific proximity labeling proteomics in challenging biological systems. Nature Communications, 16(1). https://doi.org/10.1038/s41467-025-65405-8
Edition / version
Citation
DOI
ISSN
ISBN
Language
English
Publisher
Publisher place
Classification (DDC)
570 Biology
Collections
Original object
University bibliography
Standardized keywords (GND)
Sustainable Development Goals
BibTeX
@article{Sroka2025,
doi = {10.1038/s41467-025-65405-8},
author = {Sroka, Tommy J. and Sanwald, Lea K. and Prasai, Avishek et al.},
title = {An enzymatic cascade enables sensitive and specific proximity labeling proteomics in challenging biological systems},
journal = {Nature Communications},
year = {2025},
volume = {16},
number = {1},
}