Shape changes induced by N-terminal platination of ubiquitin by cisplatin |
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Authors: | Jonathan P Williams Hazel I A Phillips Iain Campuzano Peter J Sadler |
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Institution: | 1.Department of Chemistry,University of Warwick,Coventry,UK;2.Waters Corporation,Manchester,UK |
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Abstract: | The three-dimensional conformation of a protein is an important property and plays a key role in its biological activity.
We show here that ion mobility-mass spectrometry (IM-MS) can be used to detect conformational changes in the protein ubiquitin
in the gas phase induced by reaction with the anticancer drug cisplatin. The primary adduct was ubiquitin-{Pt(NH3)2} under denaturing conditions. Up to three different conformations appear to be generated upon platination depending on the
charge state. The collision cross-sections (Ω) for each conformation indicate that the conformations of the platinated protein
are contracted in size compared with unmodified ubiquitin with generally smaller Ω values. Ion mobility-tandem MS allowed
determination of the platinum binding site without a requirement for prior Chromatographic separation. A rapid 30-min digestion
of cisplatin-modified ubiquitin with trypsin allowed the platination site to be identified as the N-terminal methionine following
low-energy collision-induced dissociation (CID) studies of the modified peptide. The data were generated using a Traveling-Wave
based ion mobility-MS approach. Such cisplatin-induced shape changes may have a significant effect on its function in vivo.
This work highlights the usefulness of the ion-mobility mass spectrometry technique for shedding new light on such protein
interactions. |
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