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Research article - Peer-reviewed, 2011

Cisplatin binds human copper chaperone Atox1 and promotes unfolding in vitro

Palm, Marie E.; Weise, Christoph F.; Lundin, Christina; Wingsle, Gunnar; Nygren, Yvonne; Björn, Erik; Naredi, Peter; Wolf-Watz, Magnus; Wittung-Stafshede, Pernilla

Abstract

Cisplatin (cisPt), Pt(NH(3))(2)Cl(2), is a cancer drug believed to kill cells via DNA binding and damage. Recent work has implied that the cellular copper (Cu) transport machinery may be involved in cisPt cell export and drug resistance. Normally, the Cu chaperone Atox1 binds Cu(I) via two cysteines and delivers the metal to metal-binding domains of ATP7B; the ATP7B domains then transfer the metal to the Golgi lumen for loading on cuproenzymes. Here, we use spectroscopic methods to test if cisPt interacts with purified Atox1 in solution in vitro. We find that cisPt binds to Atox1's metal-binding site regardless of the presence of Cu or not: When Cu is bound to Atox1, the near-UV circular dichroism signals indicate Cu-Pt interactions. From NMR data, it is evident that cisPt binds to the folded protein. CisPt-bound Atox1 is however not stable over time and the protein begins to unfold and aggregate. The reaction rates are limited by slow cisPt dechlorination. CisPt-induced unfolding of Atox1 is specific because this effect was not observed for two unrelated proteins that also bind cisPt. Our study demonstrates that Atox1 is a candidate for cisPt drug resistance: By binding to Atox1 in the cytoplasm, cisPt transport to DNA may be blocked. In agreement with this model, cell line studies demonstrate a correlation between Atox1 expression levels, and cisplatin resistance.

Published in

Proceedings of the National Academy of Sciences
2011, Volume: 108, number: 17, pages: 6951-6956
Publisher: NATL ACAD SCIENCES

    Sustainable Development Goals

    SDG3 Good health and well-being

    UKÄ Subject classification

    Forest Science

    Publication identifier

    DOI: https://doi.org/10.1073/pnas.1012899108

    Permanent link to this page (URI)

    https://res.slu.se/id/publ/59244