Analysis of Synaptic Protein–Protein Interaction by Cross-linking Mass Spectrometry
Protein–protein interactions and their dynamics define organelles’ structures and drive all aspects of cellular functions. In this chapter I describe a cross-linking mass spectrometry protocol that makes use of a newly developed MS-cleavable cross-linker to identify the protein pairs based on their proximity as defined by the length of the cross-linker. Unlike other approaches, such as immunoprecipitation-based interaction proteomics or yeast-two hybrid analysis, this protocol could characterize protein–protein interactions in a more physiological setting and allows the detection of native protein complexes contained in the mouse synaptosome.
KeywordsSynapse DSSO Cross-linking Mass Spectrometry Protein Protein interaction