Reliable identification of protein-protein interactions by crosslinking mass spectrometry

Protein-protein interactions govern most cellular pathways and processes, and multiple technologies have emerged to systematically map them. Assessing the error of interaction networks has been a challenge. Crosslinking mass spectrometry is currently widening its scope from structural analyses of purified multi-protein complexes towards systems-wide analyses of protein-protein interactions (PPIs). Using a carefully controlled large-scale analysis of Escherichia coli cell lysate, we demonstrate that false-discovery rates (FDR) for PPIs identified by crosslinking mass spectrometry can be reliably estimated. We present an interaction network comprising 590 PPIs at 1% decoy-based PPI-FDR. The structural information included in this network localises the binding site of the hitherto uncharacterised protein YacL to near the DNA exit tunnel on the RNA polymerase. Cross-linking mass spectrometry (MS) can identify protein-protein interaction (PPI) networks but assessing the reliability of these data remains challenging. To address this issue, the authors develop and validate a method to determine the false-discovery rate of PPIs identified by cross-linking MS.

Automated summary

Researchers have developed and validated a method for determining the false-discovery rate of protein-protein interactions identified by cross-linking mass spectrometry, addressing the challenge of assessing the reliability of these data.