On the Efficiency of NHS Ester Cross-Linkers for Stabilizing Integral Membrane Protein Complexes

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Author(s) Chen, Fan, Gerber, Sabina, Korkhov, Volodymyr M., Mireku, Samantha, Bucher, Monika, Locher, Kaspar P., Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title On the Efficiency of NHS Ester Cross-Linkers for Stabilizing Integral Membrane Protein Complexes
Author(s) Chen, Fan
Gerber, Sabina
Korkhov, Volodymyr M.
Mireku, Samantha
Bucher, Monika
Locher, Kaspar P.
Zenobi, Renato
Journal or Series Title Journal of the American Society for Mass Spectrometry
Volume Number 26
Issue Number 3
Start Page 493
End Page 498
ISSN 1044-0305
Publisher Springer
Publication Place New York, NY
Publication Date 2015-03
Keyword(s) Membrane protein complexes
Chemical cross-linking
Abstract We have previously presented a straightforward approach based on high-mass matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) to study membrane proteins. In addition, the stoichiometry of integral membrane protein complexes could be determined by MALDI-MS, following chemical cross-linking via glutaraldehyde. However, glutaraldehyde polymerizes in solution and reacts nonspecifically with various functional groups of proteins, limiting its usefulness for structural studies of protein complexes. Here, we investigated the capability of N-hydroxysuccinimide (NHS) esters, which react much more specifically, to cross-link membrane protein complexes such as PglK and BtuC2D2. We present clear evidence that NHS esters are capable of stabilizing membrane protein complexes in situ, in the presence of detergents such as DDM, C12E8, and LDAO. The stabilization efficiency strongly depends on the membrane protein structure (i.e, the number of primary amine groups and the distances between primary amines). A minimum number of primary amine groups is required, and the distances between primary amines govern whether a cross-linker with a specific spacer arm length is able to bridge two amine groups.
DOI 10.1007/s13361-014-1035-4
Additional Notes Received 19 August 2014, Revised 20 October 2014, Accepted 21 October 2014, Published Online 18 November 2014
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000541631
Source Database ID FORM-1417709137
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  author = "Chen, Fan and Gerber, Sabina and Korkhov, Volodymyr M. and Mireku, Samantha and Bucher, Monika and Locher, Kaspar P. and Zenobi, Renato",
  title = "{O}n the {E}fficiency of {N}{H}{S} {E}ster {C}ross-{L}inkers for {S}tabilizing {I}ntegral {M}embrane {P}rotein {C}omplexes",
  journal = "Journal of the American Society for Mass Spectrometry",
  year = 2015,
  volume = "26",
  number = "3",
  pages = "493--498",
  month = mar,

E-Citations record created: Thu, 04 Dec 2014, 16:05:44 CET