A Universal Peptide Matrix Interactomics Approach to Disclose Motif-Dependent Protein Binding.

ABSTRACT: Protein-protein interactions mediated by intrinsically disordered regions are often based on short linear motifs (SLiMs). SLiMs are implicated in signal transduction and gene regulation yet remain technically laborious and notoriously challenging to study. Here, we present an optimized method for a protein interaction screen on a peptide matrix (PRISMA) in combination with quantitative MS. The protocol was benchmarked with previously described SLiM-based protein-protein interactions using peptides derived from EGFR, SOS1, GLUT1, and CEBPB and extended to map binding partners of kinase activation loops. The detailed protocol provides practical considerations for setting up a PRISMA screen and subsequently implementing PRISMA on a liquid-handling robotic platform as a cost-effective high-thr

SUBMITTER: Ramberger E 

PROVIDER: S-EPMC8453223 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature