Mass spectrometry of membrane transporters reveals subunit stoichiometry and interactions
- 5 July 2009
- journal article
- research article
- Published by Springer Nature in Nature Methods
- Vol. 6 (8) , 585-587
- https://doi.org/10.1038/nmeth.1347
Abstract
A generally applicable approach to analyze intact membrane protein complexes by mass spectrometry is reported. This method allows subunit stoichiometry, lipid binding and the effects of post-translational modifications on complex formation to be explored. We describe a general mass spectrometry approach to determine subunit stoichiometry and lipid binding in intact membrane protein complexes. By exploring conditions for preserving interactions during transmission into the gas phase and for optimally stripping away detergent, by subjecting the complex to multiple collisions, we released the intact complex largely devoid of detergent. This enabled us to characterize both subunit stoichiometry and lipid binding in 4 membrane protein complexes.Keywords
This publication has 27 references indexed in Scilit:
- MacB ABC Transporter Is a Dimer Whose ATPase Activity and Macrolide-binding Capacity Are Regulated by the Membrane Fusion Protein MacAJournal of Biological Chemistry, 2009
- Micelles Protect Membrane Complexes from Solution to VacuumScience, 2008
- On the energy-dependence of Hoechst 33342 transport by the ABC transporter LmrABiochemical Pharmacology, 2008
- X-ray structure of EmrE supports dual topology modelProceedings of the National Academy of Sciences, 2007
- X-ray solution scattering (SAXS) combined with crystallography and computation: defining accurate macromolecular structures, conformations and assemblies in solutionQuarterly Reviews of Biophysics, 2007
- Structural Asymmetry of AcrB Trimer Suggests a Peristaltic Pump MechanismScience, 2006
- Nucleotide-Binding Sites of the Heterodimeric LmrCD ABC-Multidrug Transporter of Lactococcus lactis Are AsymmetricBiochemistry, 2005
- FUGUE: sequence-structure homology recognition using environment-specific substitution tables and structure-dependent gap penalties11Edited by B. HonigJournal of Molecular Biology, 2001
- Expression of Genes Associated with Antibiotic Extrusion in Pseudomonas aeruginosaBiochemical and Biophysical Research Communications, 1995
- Comparative Protein Modelling by Satisfaction of Spatial RestraintsJournal of Molecular Biology, 1993