Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry

Dror S Chorev; Lindsay A Baker; Di Wu; Victoria Beilsten-Edmands; Sarah L Rouse; Tzviya Zeev-Ben-Mordehai; Chimari Jiko; Firdaus Samsudin; Christoph Gerle; Syma Khalid; Alastair G Stewart; Stephen J Matthews; Kay Grünewald; Carol V Robinson

doi:10.1126/science.aau0976

Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry

Science. 2018 Nov 16;362(6416):829-834. doi: 10.1126/science.aau0976.

Authors

Dror S Chorev¹, Lindsay A Baker², Di Wu¹, Victoria Beilsten-Edmands¹, Sarah L Rouse³, Tzviya Zeev-Ben-Mordehai⁴, Chimari Jiko⁵, Firdaus Samsudin⁶, Christoph Gerle^{7

8}, Syma Khalid⁶, Alastair G Stewart^{9

10}, Stephen J Matthews³, Kay Grünewald^{2

11}, Carol V Robinson¹²

Affiliations

¹ Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK.
² Division of Structural Biology, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
³ Department of Life Sciences, Imperial College, London, South Kensington Campus, London SW7 2AZ, UK.
⁴ Division of Structural Biology, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK. carol.robinson@chem.ox.ac.uk.
⁵ Institute for Integrated Radiation and Nuclear Science, Kyoto University, Kumatori, Japan.
⁶ School of Chemistry, University of Southampton, University Road, Southampton SO17 1BJ, UK.
⁷ Institute for Protein Research, Osaka University, Suita, Osaka, Japan.
⁸ Core Research for Evolutional Science and Technology, Japan and Science and Technology Agency, Kawaguchi, Japan.
⁹ Molecular, Structural and Computational Biology Division, Victor Chang Cardiac Research Institute, Darlinghurst, NSW, Australia.
¹⁰ Faculty of Medicine, The University of New South Wales, Sydney, NSW, Australia.
¹¹ Centre of Structural Systems Biology (CSSB), Notkestr. 85, D-22607, Heinrich-Pette Institute/University of Hamburg, Hamburg, Germany.
¹² Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK. carol.robinson@chem.ox.ac.uk.

Abstract

Membrane proteins reside in lipid bilayers and are typically extracted from this environment for study, which often compromises their integrity. In this work, we ejected intact assemblies from membranes, without chemical disruption, and used mass spectrometry to define their composition. From Escherichia coli outer membranes, we identified a chaperone-porin association and lipid interactions in the β-barrel assembly machinery. We observed efflux pumps bridging inner and outer membranes, and from inner membranes we identified a pentameric pore of TonB, as well as the protein-conducting channel SecYEG in association with F₁F_O adenosine triphosphate (ATP) synthase. Intact mitochondrial membranes from Bos taurus yielded respiratory complexes and fatty acid-bound dimers of the ADP (adenosine diphosphate)/ATP translocase (ANT-1). These results highlight the importance of native membrane environments for retaining small-molecule binding, subunit interactions, and associated chaperones of the membrane proteome.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenine Nucleotide Translocator 1 / chemistry
Adenine Nucleotide Translocator 1 / metabolism*
Animals
Bacterial Outer Membrane Proteins / chemistry
Bacterial Outer Membrane Proteins / metabolism
Bacterial Proteins / chemistry
Bacterial Proteins / metabolism*
Cattle
Escherichia coli Proteins / chemistry
Escherichia coli Proteins / metabolism
Lipid Bilayers / chemistry
Lipid Bilayers / metabolism
Mass Spectrometry
Membrane Proteins / chemistry
Membrane Proteins / metabolism*
Mitochondrial Membranes / chemistry
Mitochondrial Membranes / metabolism*
Mitochondrial Proton-Translocating ATPases / chemistry
Mitochondrial Proton-Translocating ATPases / metabolism*
Molecular Chaperones / chemistry
Molecular Chaperones / metabolism*
Porins / chemistry
Porins / metabolism
Protein Conformation, beta-Strand
Proteome / chemistry
Proteome / metabolism
SEC Translocation Channels / chemistry
SEC Translocation Channels / metabolism*

Substances

Adenine Nucleotide Translocator 1
Bacterial Outer Membrane Proteins
Bacterial Proteins
Escherichia coli Proteins
Lipid Bilayers
Membrane Proteins
Molecular Chaperones
Porins
Proteome
SEC Translocation Channels
tonB protein, Bacteria
F1F0-ATP synthase
Mitochondrial Proton-Translocating ATPases

Abstract

Publication types

MeSH terms

Substances

Grants and funding