Chapter 11 Covalent Trapping of Protein Interactions in Complex Systems

摘要: Publisher Summary This chapter reviews the methodological advancements and applications aimed at study of interactomes protein complex topologies in cells tissues. The choice crosslinking reagent can strongly influence methodology outcome any experiment. A needs to be made with respect tether length nature reactive groups that mediate intended covalent crosslink. For more specialized applications, it may helpful for equipped built-in functionalities facilitate enrichment crosslinked products, allow reversal crosslinks downstream sample purification procedure, provide a means quantify crosslinks. variety strategies are currently available mapping interactions. Techniques range from genetic approaches—such as two-hybrid system (THS), synthetic lethal screens, fluorescence resonance energy transfer (FRET), quantitative interaction method—to biochemical protocols reveal composition by immunoprecipitation or affinity-tag mass spectrometry (MS). Crosslinking-based interactome studies well suited complement some purely approaches point functional interactions without establishing direct contacts proteins.