%0 Journal Article
%A Lazarewicz, Natalia
%A Le Dez, Gaëlle
%A Cerjani, Romina
%A Runeshaw, Lunelys
%A Meurer, Matthias
%A Knop, Michael
%A Wysocki, Robert
%A Rabut, Gwenaël
%T Accurate and sensitive interactome profiling using a quantitative protein-fragment complementation assay.
%J Cell reports / Methods
%V 4
%N 10
%@ 2667-2375
%C Cambridge, MA
%I Cell Press
%M DKFZ-2024-02167
%P 100880
%D 2024
%Z DKFZ-ZMBH Alliance
%X 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.
%K Protein Interaction Mapping: methods
%K Saccharomyces cerevisiae: genetics
%K Saccharomyces cerevisiae: metabolism
%K Protein Interaction Maps
%K Saccharomyces cerevisiae Proteins: metabolism
%K Saccharomyces cerevisiae Proteins: genetics
%K Luciferases: genetics
%K Luciferases: metabolism
%K CP: Systems biology (Other)
%K Cdc53 (Other)
%K Irc20 (Other)
%K Met30 (Other)
%K Nam7 (Other)
%K NanoBiT (Other)
%K Saccharomyces cerevisiae (Other)
%K Upf1 (Other)
%K budding yeast (Other)
%K interactome (Other)
%K protein-protein interaction (Other)
%K Saccharomyces cerevisiae Proteins (NLM Chemicals)
%K Luciferases (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:39437715
%R 10.1016/j.crmeth.2024.100880
%U https://inrepo02.dkfz.de/record/294334