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postgraduate thesis: Platinum on the road: the activation and transport of novel platinum anticancer drugs by the extracellulardomain of human copper transporter I (HCTR1)

TitlePlatinum on the road: the activation and transport of novel platinum anticancer drugs by the extracellulardomain of human copper transporter I (HCTR1)
Authors
Issue Date2012
PublisherThe University of Hong Kong (Pokfulam, Hong Kong)
Citation
Wang, X. [王星昊]. (2012). Platinum on the road : the activation and transport of novel platinum anticancer drugs by the extracellular domain of human copper transporter I (HCTR1). (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4819920
AbstractPlatinum-based anticancer drugs such as cisplatin, carboplatin and nedaplatin have been widely used in the chemotherapy of a variety of solid tumours for several decades. However, the development of both inherent and acquired resistance has greatly limited the efficacy of all of these drugs. Several mechanisms were proposed to explain the cellular resistance to these platinum drugs, including decreased drug accumulation. Previously, it was suggested that cisplatin enters cells via passive diffusion, followed by intracellular hydrolysis and activation prior to targeting DNA. However, recent in vivo and in vitro studies confirmed that transporters and carriers involved in copper homeostasis play important roles on the transport as well as cellular resistance to the platinum drugs. CTR1, a major plasma-membrane transporter involved in intracellular copper(I) homeostasis, was found to facilitate the uptake of several platinum drugs although the molecular mechanism remains unclear. The extracellular N-terminal domain of human CTR1 (hCTR1) with two methionine(Met)-rich and two histidine(His)-rich motifs has been proved to be essential for the uptake of both copper and platinum drugs by the transporter. In this thesis, the extracellular domain of hCTR1 (hCTR1_N, residues 1-55) was overexpressed and the role of the Met- and His-rich motifs on cisplatin binding was examined by either mutagenesis or chemical modification. Cisplatin was found to directly and rapidly bind to the Met residues of hCTR1_N by the formation of monofunctional cisplatin-thioether adducts. The kinetics of the binding process was found to correlate with the number of Met residues, indicating that all Met residues are exposed to solvents and capable for cisplatin binding. Such a non-sequence-specific binding may increase the likelihood of capturing the anticancer drug in extracellular fluid by the N-terminus of hCTR1. The effect of hCTR_N on the binding and activation of second-generation platinum anticancer drugs, e.g. carboplatin and nedaplatin, were subsequently investigated. hCTR1_N was found to significantly facilitate the activation of these platinum drugs by the formation of ring-opened monofunctional Pt-thioether species through Met residues. Although the activities of platinum drugs against hCTR1_N are significantly different, their monofunctional protein-bound species demonstrated great similarity in both structure and kinetic aspects, suggesting the uptake of these platinum drugs by hCTR1 might follow the same mechanism. The formation of active ring-opened species of carboplatin and nedaplatin by chloride/bicarbonate was observed, indicating these nucleophiles may play a critical role in the pre-activation of the drugs prior to their reaching cellular targets. Pt-thioether species were proposed as intermediates for the platination of other biomolecules. The monofunctional cisplatin adduct of hCTR1_N was proved to further transfer its active platinum species to either cysteine- or guaninecontaining biomolecules which mimic the C-ternimus of hCTR1 and DNA. Methionine residues of hCTR1 may therefore serve as key residues for the activation and transport of platinum anticancer drugs in the form of monofunctional Pt-thioether species through the pole of trimeric hCTR1 and eventually to their final target – DNA.
DegreeDoctor of Philosophy
SubjectPlatinum compounds.
Antineoplastic agents.
Copper - Physiological transport.
Dept/ProgramChemistry
Persistent Identifierhttp://hdl.handle.net/10722/188238

 

DC FieldValueLanguage
dc.contributor.authorWang, Xinghao.-
dc.contributor.author王星昊.-
dc.date.accessioned2013-08-25T02:14:21Z-
dc.date.available2013-08-25T02:14:21Z-
dc.date.issued2012-
dc.identifier.citationWang, X. [王星昊]. (2012). Platinum on the road : the activation and transport of novel platinum anticancer drugs by the extracellular domain of human copper transporter I (HCTR1). (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4819920-
dc.identifier.urihttp://hdl.handle.net/10722/188238-
dc.description.abstractPlatinum-based anticancer drugs such as cisplatin, carboplatin and nedaplatin have been widely used in the chemotherapy of a variety of solid tumours for several decades. However, the development of both inherent and acquired resistance has greatly limited the efficacy of all of these drugs. Several mechanisms were proposed to explain the cellular resistance to these platinum drugs, including decreased drug accumulation. Previously, it was suggested that cisplatin enters cells via passive diffusion, followed by intracellular hydrolysis and activation prior to targeting DNA. However, recent in vivo and in vitro studies confirmed that transporters and carriers involved in copper homeostasis play important roles on the transport as well as cellular resistance to the platinum drugs. CTR1, a major plasma-membrane transporter involved in intracellular copper(I) homeostasis, was found to facilitate the uptake of several platinum drugs although the molecular mechanism remains unclear. The extracellular N-terminal domain of human CTR1 (hCTR1) with two methionine(Met)-rich and two histidine(His)-rich motifs has been proved to be essential for the uptake of both copper and platinum drugs by the transporter. In this thesis, the extracellular domain of hCTR1 (hCTR1_N, residues 1-55) was overexpressed and the role of the Met- and His-rich motifs on cisplatin binding was examined by either mutagenesis or chemical modification. Cisplatin was found to directly and rapidly bind to the Met residues of hCTR1_N by the formation of monofunctional cisplatin-thioether adducts. The kinetics of the binding process was found to correlate with the number of Met residues, indicating that all Met residues are exposed to solvents and capable for cisplatin binding. Such a non-sequence-specific binding may increase the likelihood of capturing the anticancer drug in extracellular fluid by the N-terminus of hCTR1. The effect of hCTR_N on the binding and activation of second-generation platinum anticancer drugs, e.g. carboplatin and nedaplatin, were subsequently investigated. hCTR1_N was found to significantly facilitate the activation of these platinum drugs by the formation of ring-opened monofunctional Pt-thioether species through Met residues. Although the activities of platinum drugs against hCTR1_N are significantly different, their monofunctional protein-bound species demonstrated great similarity in both structure and kinetic aspects, suggesting the uptake of these platinum drugs by hCTR1 might follow the same mechanism. The formation of active ring-opened species of carboplatin and nedaplatin by chloride/bicarbonate was observed, indicating these nucleophiles may play a critical role in the pre-activation of the drugs prior to their reaching cellular targets. Pt-thioether species were proposed as intermediates for the platination of other biomolecules. The monofunctional cisplatin adduct of hCTR1_N was proved to further transfer its active platinum species to either cysteine- or guaninecontaining biomolecules which mimic the C-ternimus of hCTR1 and DNA. Methionine residues of hCTR1 may therefore serve as key residues for the activation and transport of platinum anticancer drugs in the form of monofunctional Pt-thioether species through the pole of trimeric hCTR1 and eventually to their final target – DNA.-
dc.languageeng-
dc.publisherThe University of Hong Kong (Pokfulam, Hong Kong)-
dc.relation.ispartofHKU Theses Online (HKUTO)-
dc.rightsThe author retains all proprietary rights, (such as patent rights) and the right to use in future works.-
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.source.urihttp://hub.hku.hk/bib/B48199205-
dc.subject.lcshPlatinum compounds.-
dc.subject.lcshAntineoplastic agents.-
dc.subject.lcshCopper - Physiological transport.-
dc.titlePlatinum on the road: the activation and transport of novel platinum anticancer drugs by the extracellulardomain of human copper transporter I (HCTR1)-
dc.typePG_Thesis-
dc.identifier.hkulb4819920-
dc.description.thesisnameDoctor of Philosophy-
dc.description.thesislevelDoctoral-
dc.description.thesisdisciplineChemistry-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.5353/th_b4819920-
dc.date.hkucongregation2012-

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