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PMID: 22021858 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mass spectrometry of intact V-type ATPases reveals bound lipids and the effects of nucleotide binding.

Science (New York, N.Y.) ·Vol. 334 ·No. 6054 ·2011-10-21 ·Pages 380-385

Zhou M, Morgner N, Barrera NP, Politis A, Isaacson SC, Matak-Vinković D, Murata T, Bernal RA, Stock D, Robinson CV

Abstract

The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.

MeSH Terms
Adenosine Triphosphatases/chemistry,metabolism Adenosine Triphosphate/metabolism Bacterial Proteins/chemistry,metabolism Binding Sites Cardiolipins/analysis,metabolism Enterococcus/enzymology Hydrolysis Hydrophobic and Hydrophilic Interactions Mass Spectrometry Membrane Lipids/analysis,metabolism Models, Molecular Phosphatidylethanolamines/analysis,metabolism Protein Conformation Protein Multimerization Protein Structure, Tertiary Protein Subunits/chemistry,metabolism Spectrometry, Mass, Electrospray Ionization Thermus thermophilus/enzymology Vacuolar Proton-Translocating ATPases/chemistry,metabolism
Chemicals
Bacterial Proteins Cardiolipins Membrane Lipids Phosphatidylethanolamines Protein Subunits phosphatidylethanolamine Adenosine Triphosphate Adenosine Triphosphatases V-type sodium ATPase, Enterococcus hirae Vacuolar Proton-Translocating ATPases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Zhou Min
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ.
Morgner Nina
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ.
Barrera Nelson P
Department of Chemistry, Lensfield Road, University of Cambridge CB2 1EW. | Department of Physiology, Pontificia Universidad Católica de Chile, Alameda 340, Santiago, Chile.
Politis Argyris
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ.
Isaacson Shoshanna C
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ.
Matak-Vinković Dijana
Department of Chemistry, Lensfield Road, University of Cambridge CB2 1EW.
Murata Takeshi
Department of Chemistry, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage, Chiba 263-8522, Japan.
Bernal Ricardo A
Department of Chemistry, University of Texas at El Paso, El Paso, Texas 79968, USA.
Stock Daniela
The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool Street, Darlinghurst NSW 2010. | Faculty of Medicine, University of New South Wales, Sydney 2052, Australia.
Robinson Carol V
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ.
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Article Info
Journal
Science (New York, N.Y.)
Abbr.
Science
ISSN
1095-9203
DOI
10.1126/science.1210148
Published
2011-10-21
Pages
380-385
Language
English
Region
United States
NLM ID
0404511
PMCID
PMC3927129
Subset
IM
Grants
Wellcome Trust · 088150 · United Kingdom
Wellcome Trust · 099141 · United Kingdom
Medical Research Council · G1000819 · United Kingdom
Corrections
CommentIn
PMID: 22021848
External Links
PubMed source DOI: 10.1126/science.1210148 PMC Full Text View journal details
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