Chemical Crosslinking in Intact Mitochondria.

Many mitochondrial proteins perform their functions as components of large, multimeric complexes. Chemical crosslinking is a powerful method to analyze protein-protein interactions within such complexes. Using membrane-permeable crosslinkers and isolated intact mitochondria, protein-protein interactions that are secluded by two mitochondrial membranes can be readily analyzed in physiologically active, isolated organelles under a variety of physiological and pathophysiological conditions. Here, we describe two methods for chemical crosslinking in intact yeast mitochondria. The first method enables the analysis of ATP-dependent remodeling of mitochondrial protein complexes while the second one allows the identification of crosslinking partners of a protein of interest.