A Cross-linking Mass Spectrometry Approach Defines Protein Interactions in Yeast Mitochondria
A. Linden, Markus Deckers, Iwan Parfentev, Ralf Pflanz, Bettina Homberg, Piotr Neumann, Ralf Ficner, Peter Rehling, Henning Urlaub
Abstract
Protein interactions in mitochondria isolated from S. cerevisiae grown on glycerol or glucose medium were analyzed by XL-MS. The non-cleavable crosslinker BS3 proved suitable for elucidating differences in protein-protein interactions under both conditions. XL-MS analysis of interprotein interactions using stable-isotope-labeled BS3 revealed certain limitations in the quantitative application of crosslinking to quite different cellular states. The results from glycerolgrown mitochondria show that Ndi1 interacts directly with CIII in an ETC supercomplex and that Min8 promotes Cox12 assembly into CIV.
Topics & Concepts
Mass spectrometryYeastChemistryMitochondrionProteomicsComputational biologyIsobaric labelingBiochemistryCell biologyChromatographyBiologyProtein mass spectrometryTandem mass spectrometryGeneMitochondrial Function and PathologyMetabolomics and Mass Spectrometry StudiesMass Spectrometry Techniques and Applications