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A universal peptide matrix interactomics approach to disclose motif dependent protein binding

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Item Type:Article
Title:A universal peptide matrix interactomics approach to disclose motif dependent protein binding
Creators Name:Ramberger, E. and Suarez-Artiles, L. and Perez-Hernandez, D. and Haji, M. and Popp, O. and Reimer, U. and Leutz, A. and Dittmar, G. and Mertins, P.
Abstract:Protein-protein interactions (PPIs) mediated by intrinsically disordered regions (IDRs) are often based on short linear motifs (SLiM). 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 mass spectrometry. The protocol was benchmarked with previously described SLiM based PPIs 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-throughput method. Optimized PRISMA can be universally applied to systematically study SLiM based interactions and associated post translational modifications (PTMs) or mutations to advance our understanding of the largely uncharacterized interactomes of intrinsically disordered protein regions.
Keywords:Amino Acid Motifs, HeLa Cells, Peptides, Point Mutation, Protein Binding, Protein Interaction Domains and Motifs, Post-Translational Protein Processing, Proteomics / Methods
Source:Molecular & Cellular Proteomics
Publisher:American Society for Biochemistry and Molecular Biology
Page Range:100135
Date:13 August 2021
Official Publication:https://doi.org/10.1016/j.mcpro.2021.100135
PubMed:View item in PubMed

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