Cisplatin affects the conformation of apo form, not holo form, of BRCA1 RING finger domain and confers thermal stability.
The breast cancer suppressor protein 1 (BRCA1) has been shown to participate in genomic integrity maintenance. Preclinical and clinical studies have recently revealed that the inactivation of BRCA1 in cancer cells leads to chemosensitivity. Approaching the BRCA1 RING protein as a potentially molecular target for a platinum-based drug might be of interest in cancer therapy. In the present study, the in vitro platination of the BRCA1 RING protein by the anticancer drug cisplatin was observed. The protein contained a preformed structure in the apo form with structural changes and resistance to limited proteolysis after Zn2+ binding. SDS-PAGE and mass-spectrometric analyses revealed that cisplatin preferentially formed monofunctional and bifunctional BRCA1 adducts. Tandem mass spectrometry (MS/MS) of the 656.29(2+) ion indicated that the ion arose from [Pt(NH3)2(OH)]+ bound to the BRCA1 peptide (111)ENNSPEHLK(119). The product-ion spectrum revealed the Pt-binding site on His117. Circular dichroism showed that the apo form, not holo form, of BRCA1 underwent more folded structural rearrangement upon cisplatin binding. Cisplatin-bound protein exhibited an enhanced thermostability by 13 degrees , resulting from the favorably intermolecular cross-links driven by the free energy. Our findings demonstrated the first conformational and thermal evidences for a direct binding of cisplatin to the BRCA1 RING domain and could raise a possibility of selectively targeted treatment of cancer with less toxicity or improved response to conventional regimens.
Atipairin
Apichart
A
Laboratory of Pharmaceutical Biotechnology, Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand.
Canyuk
Bhutorn
B
Ratanaphan
Adisorn
A
D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000494:pharmacology; D019313:BRCA1 Protein / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D002945:Cisplatin / Q000378:metabolism / Q000494:pharmacology*; D019281:Dimerization; D004355:Drug Stability; D006801:Humans; D008969:Molecular Sequence Data; D054829:RING Finger Domains / Q000187:drug effects*; D013696:Temperature
D000970:Antineoplastic Agents; D019313:BRCA1 Protein; D002945:Cisplatin