A quantitative intracellular peptide-binding assay reveals recognition determinants and context dependence of short linear motifs.

ABSTRACT: Transient protein-protein interactions play key roles in controlling dynamic cellular responses. Many examples involve globular protein domains that bind to peptide sequences known as short linear motifs (SLiMs), which are enriched in intrinsically disordered regions of proteins. Here we describe a novel functional assay for measuring SLiM binding, called systematic intracellular motif-binding analysis (SIMBA). In this method, binding of a foreign globular domain to its cognate SLiM peptide allows yeast cells to proliferate by blocking a growth arrest signal. A high-throughput application of the SIMBA method involving competitive growth and deep sequencing provides rapid quantification of the relative binding strength for thousands of SLiM sequence variants and a comprehensive interrogatio

SUBMITTER: Subbanna MS 

PROVIDER: S-EPMC11879687 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature