Article Dans Une Revue Cell Reports Methods Année : 2024

Accurate and sensitive interactome profiling using a quantitative protein-fragment complementation assay

Résumé

An accurate description of protein-protein interaction (PPI) networks is key to understanding the molecular mechanisms underlying cellular systems. Here, we constructed genome-wide libraries of yeast strains to systematically probe protein-protein interactions using NanoLuc Binary Technology (NanoBiT), a quantitative protein-fragment complementation assay (PCA) based on the NanoLuc luciferase. By investigating an array of well-documented PPIs as well as the interactome of four proteins with varying levels of characterization-including the well-studied nonsense-mediated mRNA decay (NMD) regulator Upf1 and the SCF complex subunits Cdc53 and Met30-we demonstrate that ratiometric NanoBiT measurements enable highly precise and sensitive mapping of PPIs. This work provides a foundation for employing NanoBiT in the assembly of more comprehensive and accurate protein interaction maps as well as in their functional investigation.
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hal-04767463 , version 1 (05-11-2024)

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Natalia Lazarewicz, Gaëlle Le Dez, Romina Cerjani, Lunelys Runeshaw, Matthias Meurer, et al.. Accurate and sensitive interactome profiling using a quantitative protein-fragment complementation assay. Cell Reports Methods, 2024, 4 (10), pp.100880. ⟨10.1016/j.crmeth.2024.100880⟩. ⟨hal-04767463⟩
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