Characterization of protein complexes using chemical cross-linking coupled electrospray mass spectrometry

Abstract

Identification and characterization of large protein complexes is a mainstay of biochemical toolboxes. Utilization of cross-linking chemicals can facilitate the capture and identification of transient or weak interactions of a transient nature. Here we describe a detailed methodology for cell culture based proteomic approach. We describe the generation of cells stably expressing green fluorescent protein (GFP)- tagged proteins under the tetracycline-inducible promoter and subsequent proteomic analysis of GFP-interacting proteins. We include a list of proteins that were identified as interactors of GFP.