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PMID: 19578383 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mass spectrometry of membrane transporters reveals subunit stoichiometry and interactions.

Nature methods ·Vol. 6 ·No. 8 ·2009-08-00 ·Pages 585-7

Barrera NP, Isaacson SC, Zhou M, Bavro VN, Welch A, Schaedler TA, Seeger MA, Miguel RN, Korkhov VM, van Veen HW, Venter H, Walmsley AR, Tate CG, Robinson CV

Abstract

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.

MeSH Terms
Membrane Transport Proteins/chemistry Multiprotein Complexes/chemistry Protein Interaction Mapping Protein Subunits/chemistry Proteomics/methods Spectrometry, Mass, Electrospray Ionization/methods
Chemicals
Membrane Transport Proteins Multiprotein Complexes Protein Subunits
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Barrera Nelson P
Department of Chemistry, University of Cambridge, Cambridge, UK.
Isaacson Shoshanna C
Zhou Min
Bavro Vassiliy N
Welch Alex
Schaedler Theresia A
Seeger Markus A
Miguel Ricardo Núñez
Korkhov Vladimir M
van Veen Hendrik W
Venter Henrietta
Walmsley Adrian R
Tate Christopher G
Robinson Carol V
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.1347
Published
2009-08-00
Epub
2009-00-05
Pages
585-7
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC4066579
Subset
IM
Grants
Wellcome Trust · 088150 · United Kingdom
Biotechnology and Biological Sciences Research Council · BB/F008333/1 · United Kingdom
Medical Research Council · United Kingdom
External Links
PubMed source DOI: 10.1038/nmeth.1347 PMC Full Text View journal details
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