title: A comparative study on the interactions of human copper chaperone Cox17 with anticancer organoruthenium(II) complexes and cisplatin by mass spectrometry.
abstract: Herein we report investigation of the interactions between anticancer organoruthenium complexes, [(η(6)-arene)Ru(en)(Cl)]PF6 (en=ethylenediamine, arene=p-cymene (1) or biphenyl (2)), and the human copper chaperone protein Cox17 by mass spectrometry with cisplatin as a reference. The electrospray ionization mass spectrometry (ESI-MS) results indicate much weaker binding of the ruthenium complexes than that of cisplatin to apo-Cox172s-s, the functional state of Cox17. Up to tetra-platinated Cox17 adducts were identified while only mono-ruthenated and a little amount of di-ruthenated Cox17 adducts were detected even for the reactions with 10-fold excess of the Ru complexes. However, ESI-MS analysis coupled with liquid chromatography of tryptic digests of metalated proteins identified only three platination sites as Met4, Cys27 and His47 residues, possibly due to the lower abundance or facile dissociation of Pt bindings at other sites. Complexes 1 and 2 were found to bind to the same three residues with Met4 as the major site. Inductively coupled plasma mass spectrometry results revealed that ~7mol Pt binding to 1mol apo-Cox172s-s molecules, compared to only 0.17 (1) and 0.10 (2) mol Ru to 1mol apo-Cox172s-s. This is in line with the circular dichroism results that much larger unfolding extent of α-helix of apo-Cox172s-s was observed upon cisplatin binding than that upon organoruthenium bindings. These results collectively indicate that Cox17 might not participate in the action of these anticancer organoruthenium complexes, and further verify the distinct anticancer mechanism of the organoruthenium(II) complexes from cisplatin.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China; School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing 100083, PR China.
authors.forename: Lijie
authors.initials: L
authors.lastname: Li
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China. Electronic address: wukui@iccas.ac.cn.
authors.forename: Kui
authors.initials: K
authors.lastname: Wu
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, PR China.
authors.forename: Xuelei
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, PR China.
authors.forename: Linhong
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Qun
authors.initials: Q
authors.lastname: Luo
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, PR China.
authors.forename: Yangzhong
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing 100083, PR China.
authors.forename: Qingwu
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, National Centre for Mass Spectrometry in Beijing, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China. Electronic address: fuyi.wang@iccas.ac.cn.
authors.forename: Fuyi
authors.initials: F
authors.lastname: Wang
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D002945:Cisplatin / Q000737:chemistry*; D000079922:Copper Transport Proteins; D006801:Humans; D013058:Mass Spectrometry; D016566:Organoselenium Compounds / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; C491006:COX17 protein, human; D002352:Carrier Proteins; D000079922:Copper Transport Proteins; D016566:Organoselenium Compounds; D002945:Cisplatin
keywords: Anticancer agents; Cisplatin; Cox17; LC-ESI-MS; Organoruthenium complexes