Protein interaction landscapes revealed by advanced in vivo cross-linking–mass spectrometry

Abstract

Significance Blueprints of in-cell protein interaction landscapes are essential for our understanding of cellular structures and functions, which have been challenging to study at the systems level. Cross-linking–mass spectrometry (XL-MS) represents a high-throughput method for global profiling of interaction networks and can determine the identity and connectivity of native PPIs simultaneously without cell engineering. While in vivo XL-MS experiments are feasible, in-depth analyses remain difficult due to technical limitations on sample preparation. Here, we have developed a new Alkyne-A-DSBSO–based in vivo XL-MS platform that enabled us to obtain the most comprehensive PPI maps in cells. This approach can be adopted for proteome-wide XL-MS studies in any organisms and sample origins, thus advancing interactome biology beyond proteome identity and abundance.

Keywords

Affiliated Institutions

• University of California, Irvine US
• Gladstone Institutes US
• University of California, San Francisco US
• Beijing Proteome Research Center CN

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