Proximity labeling and identification of endogenous client proteins recruited to Y15-based artificial granules tethering a bait protein.

Autor: Hashimoto M; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan., Miki T; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.; Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, Tokyo, Japan., Niwa T; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.; Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan., Mihara H; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
Jazyk: angličtina
Zdroj: Journal of peptide science : an official publication of the European Peptide Society [J Pept Sci] 2024 Feb; Vol. 30 (2), pp. e3536. Date of Electronic Publication: 2023 Aug 14.
DOI: 10.1002/psc.3536
Abstrakt: Protein clustering is a ubiquitous event in diverse cellular processes. Self-association of proteins triggers recruitment of downstream proteins to regulate cellular signaling. To investigate the interactions in detail, chemical biology tools to identify proteins recruited to defined assemblies are required. Here, we exploit an identification of proteins recruited in artificial granules (IPRAG) platform that combines intracellular Y15-based supramolecule construction with a proximity labeling method. We validated the IPRAG tool using Nck1 as a target bait protein. We constructed Nck1-tethering granules, labeled the recruited proteins with biotin, and analyzed them by LC-MS/MS. As a result, we successfully identified proteins that directly or indirectly interact with Nck1.
(© 2023 The Authors. Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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