Abstract
Cisplatin, carboplatin, and oxaliplatin are widely used anticancer drugs. Their efficacy is strongly reduced by development of cell resistance. Down-regulation of CTR1 and up-regulation of the Cu-ATPases, ATP7A and ATP7B, have been associated to augmented drug resistance. To gain information on translocation of Pt drugs by human Cu-ATPases, we performed electrical measurements on the COS-1 cell microsomal fraction, enriched with recombinant ATP7A, ATP7B, and selected mutants, and adsorbed on a solid supported membrane. The experimental results indicate that Pt drugs activate Cu-ATPases and undergo ATP-dependent translocation in a fashion similar to that of Cu. We then used NMR spectroscopy and ESI-MS to determine the binding mode of these drugs to the first N-terminal metal-binding domain of ATP7A (Mnk1).
Keywords:
ATPases; NMR spectroscopy; charge measurements; copper; platinum drugs.
Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / chemistry
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism*
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Adenosine Triphosphate / metabolism
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Amino Acid Motifs
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Animals
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Antineoplastic Agents / chemistry*
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Antineoplastic Agents / metabolism
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Antineoplastic Agents / toxicity
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COS Cells
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Cation Transport Proteins / chemistry
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Cation Transport Proteins / genetics
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Cation Transport Proteins / metabolism*
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Chlorocebus aethiops
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Cisplatin / chemistry*
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Cisplatin / metabolism
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Cisplatin / toxicity
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Copper / chemistry
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Copper / metabolism
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Copper Transporter 1
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Copper-Transporting ATPases
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Down-Regulation / drug effects
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Drug Resistance, Neoplasm
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Humans
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Intracellular Signaling Peptides and Proteins / chemistry
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Intracellular Signaling Peptides and Proteins / metabolism
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Magnetic Resonance Spectroscopy
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Microsomes / metabolism
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Mutagenesis, Site-Directed
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Organoplatinum Compounds / chemistry*
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Organoplatinum Compounds / metabolism
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Organoplatinum Compounds / toxicity
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Oxaliplatin
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Protein Binding
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Protein Serine-Threonine Kinases / chemistry
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Protein Serine-Threonine Kinases / metabolism
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Spectrometry, Mass, Electrospray Ionization
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Up-Regulation / drug effects
Substances
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Antineoplastic Agents
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Cation Transport Proteins
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Copper Transporter 1
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Intracellular Signaling Peptides and Proteins
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Organoplatinum Compounds
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Recombinant Proteins
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SLC31A1 protein, human
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Oxaliplatin
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Copper
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Adenosine Triphosphate
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MKNK1 protein, human
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Protein Serine-Threonine Kinases
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Adenosine Triphosphatases
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ATP7A protein, human
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ATP7B protein, human
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Copper-Transporting ATPases
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Cisplatin