A toolbox for systematic discovery of stable and transient protein interactors in baker's yeast.

ABSTRACT: Identification of both stable and transient interactions is essential for understanding protein function and regulation. While assessing stable interactions is more straightforward, capturing transient ones is challenging. In recent years, sophisticated tools have emerged to improve transient interactor discovery, with many harnessing the power of evolved biotin ligases for proximity labelling. However, biotinylation-based methods have lagged behind in the model eukaryote, Saccharomyces cerevisiae, possibly due to the presence of several abundant, endogenously biotinylated proteins. In this study, we optimised robust biotin-ligation methodologies in yeast and increased their sensitivity by creating a bespoke technique for downregulating endogenous biotinylation, which we term ABOLISH (Auxi

SUBMITTER: Fenech EJ 

PROVIDER: S-EPMC9912024 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature