Li, Huilin, Snelling, Jonathon R., Barrow, Mark P., Scrivens, James H., Sadler, P. J. and O’Connor, Peter B. (2014) Mass spectrometric strategies to improve the identification of Pt(II)-modification sites on peptides and proteins. Journal of The American Society for Mass Spectrometry, 25 (7). pp. 1217-1227. doi:10.1007/s13361-014-0877-0 ISSN 1044-0305.
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Abstract
To further explore the binding chemistry of cisplatin (cis-Pt(NH3)2Cl2) to peptides and also establish mass spectrometry (MS) strategies to quickly assign the platinum-binding sites, a series of peptides with potential cisplatin binding sites (Met(S), His(N), Cys(S), disulfide, carboxyl groups of Asp and Glu, and amine groups of Arg and Lys, were reacted with cisplatin, then analyzed by electron capture dissociation (ECD) in a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS). Radical-mediated side-chain losses from the charge-reduced Pt-binding species (such as CH3S(•) or CH3SH from Met, SH(•) from Cys, CO2 from Glu or Asp, and NH2(•) from amine groups) were found to be characteristic indicators for rapid and unambiguous localization of the Pt-binding sites to certain amino acid residues. The method was then successfully applied to interpret the top-down ECD spectrum of an inter-chain Pt-crosslinked insulin dimer, insulin + Pt(NH3)2 + insulin (>10 kDa). In addition, ion mobility MS shows that Pt binds to multiple sites in Substance P, generating multiple conformers, which can be partially localized by collisionally activated dissociation (CAD). Platinum(II) (Pt(II)) was found to coordinate to amine groups of Arg and Lys, but not to disulfide bonds under the conditions used. The coordination of Pt to Arg or Lys appears to arise from the migration of Pt(II) from Met(S) as shown by monitoring the reaction products at different pH values by ECD. No direct binding of cisplatin to amine groups was observed at pH 3 ~ 10 unless Met residues were present in the sequence, but noncovalent interactions between cisplatin hydrolysis and amination [Pt(NH3)4](2+) products and these peptides were found regardless of pH.
| Item Type: | Journal Article |
|---|---|
| Subjects: | Q Science > QC Physics |
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
| Library of Congress Subject Headings (LCSH): | Mass spectrometry, Ion mobility spectroscopy, Post-translational modification |
| Journal or Publication Title: | Journal of The American Society for Mass Spectrometry |
| Publisher: | Springer New York LLC |
| ISSN: | 1044-0305 |
| Official Date: | July 2014 |
| Dates: | Date Event July 2014 Published 21 May 2014 Available 23 February 2014 Accepted 21 December 2012 Submitted |
| Volume: | 25 |
| Number: | 7 |
| Number of Pages: | 11 |
| Page Range: | pp. 1217-1227 |
| DOI: | 10.1007/s13361-014-0877-0 |
| Status: | Peer Reviewed |
| Publication Status: | Published |
| Access rights to Published version: | Restricted or Subscription Access |
| Funder: | University of Warwick Postgraduate Research Scholarship, Engineering and Physical Sciences Research Council (EPSRC), Royal Society of Chemistry, National Institutes of Health (NIH), European Research Council (ERC) |
| Grant number: | NIH/NIGMS-R01GM078293 (NIH), EP/F034210/1 (EPSRC), BP/G006792 (EPSRC), 247450 (ERC) |
| URI: | https://wrap.warwick.ac.uk/70124/ |
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