title: Platination of the copper transporter ATP7A involved in anticancer drug resistance.
abstract: The clinical efficacy of the widely used anticancer drug cisplatin is severely limited by the emergence of resistance. This is related to the drug binding to proteins such as the copper influx transporter Ctr1, the copper chaperone Atox1, and the copper pumps ATP7A and ATP7B. While the binding modes of cisplatin to the first two proteins are known, the structural determinants of platinated ATP7A/ATP7B are lacking. Here we investigate the interaction of cisplatin with the first soluble domain of ATP7A. First, we establish by ESI-MS and (1)H, (13)C, and (15)N NMR that, in solution, the adduct is a monomer in which the sulfur atoms of residues Cys19 and Cys22 are cis-coordinated to the [Pt(NH3)2](2+) moiety. Then, we carry out hybrid Car-Parrinello QM/MM simulations and computational spectroscopy calculations on a model adduct based on the NMR structure of the apo protein and featuring the experimentally determined binding mode of the metal ion. These calculations show quantitative agreement with CD spectra and (1)H, (13)C, and (15)N NMR chemical shifts, thus providing a quantitative molecular view of the 3D binding mode of cisplatin to ATP7A. Importantly, the same comparison rules out a variety of alternative models with different coordination modes, that we explored to test the robustness of the computational approach. Using this combined in silico-in vitro approach we provide here for the first time a quantitative 3D atomic view of the platinum binding to the first soluble domain of ATP7A.
authors.affiliation: Computational Biophysics, German Research School for Simulation Sciences, 52425 Jülich, Germany. p.carloni@grs-sim.de.
authors.forename: Vania
authors.initials: V
authors.lastname: Calandrini
authors.forename: Fabio
authors.initials: F
authors.lastname: Arnesano
authors.forename: Angela
authors.initials: A
authors.lastname: Galliani
authors.forename: Trung Hai
authors.initials: TH
authors.lastname: Nguyen
authors.forename: Emiliano
authors.initials: E
authors.lastname: Ippoliti
authors.forename: Paolo
authors.initials: P
authors.lastname: Carloni
authors.forename: Giovanni
authors.initials: G
authors.lastname: Natile
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000737:chemistry*; D027682:Cation Transport Proteins / Q000737:chemistry*; D002942:Circular Dichroism; D002945:Cisplatin / Q000737:chemistry*; D003198:Computer Simulation; D003300:Copper; D000073840:Copper-Transporting ATPases; D019008:Drug Resistance, Neoplasm; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000970:Antineoplastic Agents; D027682:Cation Transport Proteins; D003300:Copper; D000251:Adenosine Triphosphatases; C081184:ATP7A protein, human; D000073840:Copper-Transporting ATPases; D002945:Cisplatin