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. 2016 Nov 22;6:37581. doi: 10.1038/srep37581

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(A) His-PBD_326-603 (4 μg) was treated with 2.5 mM TQ, 250 μM Poloxin (2.5 mM precipitated), or DMSO (negative control) for approximately 2 hours. Samples were submitted to MALDI-TOF analysis. The full spectra showing the 3 forms of the protein (+1 peak around 36.9 kDa, +2 peak around 18.5 kDa, and the +3 peak around 12 kDa), are shown. The +1 form was zoomed in to better evaluate the peak displacement of the PBD caused by alkylation by TQ and Poloxin. (B) Alkylation sites detected by LC-MS/MS on tryptic digests of alkylated PBD. Residues observed to be alkylated by TQ, Poloxin and Cpd 161 are indicated by colored circles. In each case, the grey line indicates the sequence covered by the peptides observed in the analysis, with percentages of PBD sequence on the right.