| Summary: | Protein-protein interactions are at the core of all cellular functions and dynamic alterations in protein interactions regulate cellular signaling. In the last decade, mass spectrometry-based proteomics has delivered unprecedented insights into human protein interaction networks. Affinity purification-mass spectrometry has been extensively employed for focused and high-throughput studies of steady state protein-protein interactions. Future challenges remain in mapping transient protein interactions after cellular perturbations as well as in resolving the spatial organization of protein interaction networks. Affinity purification-mass spectrometry can be combined with quantitative proteomics approaches to determine the relative abundance of purified proteins in different conditions, thereby enabling the identification of transient protein interactions. In addition to affinity purification, methods based on protein co-fractionation have been combined with quantitative mass spectrometry to map transient protein interactions during cellular signaling. More recently, approaches based on proximity tagging that preserve the spatial dimension of protein interaction networks have been introduced. Here, we provide an overview of mass spectrometry-based methods for analyzing protein-protein interactions with a focus on approaches that aim to dissect the temporal and spatial aspects of protein interaction networks.
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