High-Mass Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry of Integral Membrane Proteins and Their Complexes

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Author(s) Chen, Fan, Gerber, Sabina, Heuser, Katrin, Korkhov, Vladimir M., Lizak, Christian, Mireku, Samantha, Locher, Kaspar P., Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title High-Mass Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry of Integral Membrane Proteins and Their Complexes
Author(s) Chen, Fan
Gerber, Sabina
Heuser, Katrin
Korkhov, Vladimir M.
Lizak, Christian
Mireku, Samantha
Locher, Kaspar P.
Zenobi, Renato
Journal or Series Title Analytical chemistry
Volume Number 85
Issue Number 7
Start Page 3483
End Page 3488
ISSN 0003-2700
Publisher American Chemical Society
Publication Place Washington, DC
Publication Date 2013-04-02
Abstract Analyzing purified membrane proteins and membrane protein complexes by mass spectrometry has been notoriously challenging and required highly specialized buffer conditions, sample preparation methods, and apparatus. Here we show that a standard matrix-assisted laser desorption/ionization (MALDI) protocol, if used in combination with a high-mass detector, allows straightforward mass spectrometric measurements of integral membrane proteins and their complexes, directly following purification in detergent solution. Molecular weights can be determined precisely (mass error <= 0.1%) such that high-mass MALDI-MS was able to identify the site for N-linked glycosylation of the eukaryotic multidrug ABC transporter Cdrlp without special purification steps, which is impossible by any other current approach. After chemical cross-linking with glutaraldehyde in the presence of detergent micelles, the subunit stoichiometries of a series of integral membrane protein complexes, including the homomeric PglK and the heteromeric BtuCD as well as BtuCDF, were unambiguously resolved. This thus adds a valuable tool for biophysical characterization of integral membrane proteins.
DOI 10.1021/ac4000943
Additional Notes Received 10 January 2013, Accepted 6 March 2013, Published online 6 March 2013
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03652
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NEBIS System Number 000005465
Source Database ID WOS-000317173200005
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  author = "Chen, Fan and Gerber, Sabina and Heuser, Katrin and Korkhov, Vladimir M. and Lizak, Christian and Mireku, Samantha and Locher, Kaspar P. and Zenobi, Renato",
  title = "{H}igh-{M}ass {M}atrix-{A}ssisted {L}aser {D}esorption {I}onization-{M}ass {S}pectrometry of {I}ntegral {M}embrane {P}roteins and {T}heir {C}omplexes",
  journal = "Analytical chemistry",
  year = 2013,
  volume = "85",
  number = "7",
  pages = "3483--3488",
  month = apr,

E-Citations record created: Tue, 21 May 2013, 08:17:12 CET