Reactivity and applications of new amine reactive cross-linkers for mass spectrometric detection of protein-protein complexes

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Author(s) Bich, Claudia, Maedler, Stefanie, Chiesa, Katja, DeGiacomo, Fabio, Bogliotti, Nicolas, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Reactivity and applications of new amine reactive cross-linkers for mass spectrometric detection of protein-protein complexes
Author(s) Bich, Claudia
Maedler, Stefanie
Chiesa, Katja
DeGiacomo, Fabio
Bogliotti, Nicolas
Zenobi, Renato
Journal or Series Title Analytical chemistry
Volume Number 82
Issue Number 1
Start Page 172
End Page 179
ISSN 0003-2700
1520-6882
Publisher American Chemical Society
Publication Place Washington, D.C
Publication Date 2010
Abstract Chemical cross-linking of proteins permits the stabilization of noncovalent complexes through introduction of covalent bonds. A crucial challenge is to find the fastest and most efficient cross-linkers in order to minimize reaction times and to handle delicate complexes. New cross-linkers were synthesized by introducing N-hydroxyphthalimide, hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole as leaving groups instead of the commonly used N-hydroxysuccimidyl moiety. With the use of matrix-assisted laser desorption ionization (MALDI) mass spectrometry, these new cross-linkers were then compared with the commercially available disuccinimidyl suberate (DSS) for covalent stabilization of the gluthatione-S-transferase (GST) dimer and of an antibody-antigen complex. They showed a better efficiency, generated about 30% more cross-linked complex, and reacted about 10 times faster than DSS. The reaction with the GST dimer was utilized to get information about their reaction efficiency and kinetics. Their ability to stabilize only specific protein complexes was verified by incubating them with a mixture of the proteins GST and ubiquitin. Finally, the cross-linkers were incubated with synthetic peptides to study the selectivity of the binding with various amino acid side chains. Not only lysine but also tyrosine was found to react with the newly synthesized cross-linker containing 1-hydroxy-7-azabenzotriazole as the reactive group.
DOI 10.1021/ac901651r
Additional Notes Received 24 July 2009, Accepted 17 November 2009, Publication Date (Web) 8 December 2009
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000005465
Source Database ID WOS-000273265700032
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@article{Bch2010,
  author = "Bich, Claudia and Maedler, Stefanie and Chiesa, Katja and DeGiacomo, Fabio and Bogliotti, Nicolas and Zenobi, Renato",
  title = "{R}eactivity and applications of new amine reactive cross-linkers for mass spectrometric detection of protein-protein complexes",
  journal = "Analytical chemistry",
  year = 2010,
  volume = "82",
  number = "1",
  pages = "172--179",
}


E-Citations record created: Tue, 20 Apr 2010, 06:26:34 CET