title: Chemical Crosslinking in Intact Mitochondria.
abstract: 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.
authors.affiliation: Biomedical Center-Physiological Chemistry, LMU Munich, Großhadernerstr. 9, 82152, Martinsried, Germany.
authors.forename: Rupa
authors.initials: R
authors.lastname: Banerjee
authors.affiliation: Biomedical Center-Physiological Chemistry, LMU Munich, Großhadernerstr. 9, 82152, Martinsried, Germany.
authors.forename: Umut
authors.initials: U
authors.lastname: Günsel
authors.affiliation: Biomedical Center-Physiological Chemistry, LMU Munich, Großhadernerstr. 9, 82152, Martinsried, Germany. Dejana.Mokranjac@med.uni-muenchen.de.
authors.forename: Dejana
authors.initials: D
authors.lastname: Mokranjac
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D003432:Cross-Linking Reagents; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D003432:Cross-Linking Reagents; D024101:Mitochondrial Proteins; D046912:Multiprotein Complexes; D000255:Adenosine Triphosphate
keywords: Crosslinking; Mitochondria; Protein complex; Protein-protein interactions; Remodeling of protein complexes; TIM23 complex