Analytical and Bioanalytical Chemistry

, Volume 402, Issue 8, pp 2655–2662 | Cite as

Fragmentation methods on the balance: unambiguous top–down mass spectrometric characterization of oxaliplatin–ubiquitin binding sites

  • Samuel M. Meier
  • Yury O. Tsybin
  • Paul J. Dyson
  • Bernhard K. Keppler
  • Christian G. HartingerEmail author
Original Paper


The interaction between oxaliplatin and the model protein ubiquitin (Ub) was investigated in a top–down approach by means of high-resolution electrospray ionization mass spectrometry (ESI-MS) using diverse tandem mass spectrometric (MS/MS) techniques, including collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD). To the best of our knowledge, this is the first time that metallodrug–protein adducts were analyzed for the metal-binding site by ETD-MS/MS, which outperformed both CID and HCD in terms of number of identified metallated peptide fragments in the mass spectra and the localization of the binding sites. Only ETD allowed the simultaneous and exact determination of Met1 and His68 residues as binding partners for oxaliplatin. CID-MS/MS experiments were carried out on orbitrap and ion cyclotron resonance (ICR)-FT mass spectrometers and both instruments yielded similar results with respect to number of metallated fragments and the localization of the binding sites. A comparison of the protein secondary structure with the intensities of peptide fragments generated by collisional activation of the [Ub + Pt-(chxn)] adduct [chxn = (1R,2R)-cyclohexanediamine] revealed a correlation with cleavages in solution phase random coil areas, indicating that the N-terminal β-hairpin and α-helix structures are retained in the gas phase.


CID, HCD and ETD were used to determine the binding site of the anticancer agent oxaliplatin on ubiquitin in a top-down approach


Anticancer metallodrugs Tandem mass spectrometry Electron transfer dissociation Oxaliplatin Ubiquitin 



The authors are indebted to the Austrian Science Fund (FWF; I496-B11), the Hochschuljubiläumsstiftung Vienna and COST D39 and CM0902. We would like to thank Dr. Yue Xuan (Thermo Scientific) and Dr. Jens Fuchser (Bruker Daltonics) for assistance during measurements on the orbitrap FT MS and FT-ICR MS instruments, respectively.

Supplementary material

216_2011_5523_MOESM1_ESM.pdf (598 kb)
Specifications on the detected platinated peptide fragments including accurate mass, exact mass, mass accuracy, relative abundance, resolution, and number. (PDF 597 kb)


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Samuel M. Meier
    • 1
    • 2
  • Yury O. Tsybin
    • 3
  • Paul J. Dyson
    • 3
  • Bernhard K. Keppler
    • 1
    • 2
  • Christian G. Hartinger
    • 1
    • 2
    • 3
    Email author
  1. 1.Institute of Inorganic ChemistryUniversity of ViennaViennaAustria
  2. 2.Research Platform “Translational Cancer Therapy Research”University of ViennaViennaAustria
  3. 3.Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de Lausanne (EPFL)LausanneSwitzerland

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