title: Crystals of Na(+)/K(+)-ATPase with bound cisplatin.
abstract: Cisplatin is the most widely used chemotherapeutics for cancer treatment, however, its administration is connected to inevitable adverse effects. Previous studies suggested that cisplatin is able to inhibit Na(+)/K(+)-ATPase (NKA), the enzyme responsible for maintaining electrochemical potential and sodium gradient across the plasma membrane. Here we report a crystallographic analysis of cisplatin bound to NKA in the ouabain bound E2P form. Despite a moderate resolution (7.4 Å and 7.9 Å), the anomalous scattering from platinum and a model representation from a recently published structure enabled localization of seven cisplatin binding sites by anomalous difference Fourier maps. Comparison with NKA structures in the E1P conformation suggested two possible inhibitory mechanisms for cisplatin. Binding to Met151 can block the N-terminal pathway for transported cations, while binding to Met171 can hinder the interaction of cytoplasmic domains during the catalytic cycle.
authors.affiliation: Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, 17. listopadu 12, 771 46 Olomouc, Czech Republic; Aarhus University, Department of Molecular Biology and Genetics, Gustav Wieds Vej 10C, DK-8000 Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, DK-8000 Aarhus, Denmark. Electronic address: miroslavhuliciak@gmail.com.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Huliciak
authors.affiliation: Aarhus University, Department of Molecular Biology and Genetics, Gustav Wieds Vej 10C, DK-8000 Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, DK-8000 Aarhus, Denmark. Electronic address: linda.reinhard@ki.se.
authors.forename: Linda
authors.initials: L
authors.lastname: Reinhard
authors.affiliation: Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, DK-8000 Aarhus, Denmark; Department of Biomedicine, Aarhus University, Ole Worms Alle 6, DK-8000 Aarhus, Denmark. Electronic address: mette.laursen@embl-hamburg.de.
authors.forename: Mette
authors.initials: M
authors.lastname: Laursen
authors.affiliation: Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, DK-8000 Aarhus, Denmark; Department of Biomedicine, Aarhus University, Ole Worms Alle 6, DK-8000 Aarhus, Denmark. Electronic address: nf@biomed.au.dk.
authors.forename: Natalya
authors.initials: N
authors.lastname: Fedosova
authors.affiliation: Aarhus University, Department of Molecular Biology and Genetics, Gustav Wieds Vej 10C, DK-8000 Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, DK-8000 Aarhus, Denmark. Electronic address: pn@mb.au.dk.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
authors.affiliation: Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, 17. listopadu 12, 771 46 Olomouc, Czech Republic. Electronic address: mkubala@prfnw.upol.cz.
authors.forename: Martin
authors.initials: M
authors.lastname: Kubala
mesh_terms: D002945:Cisplatin / Q000009:adverse effects / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D008715:Methionine / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D010042:Ouabain / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D010042:Ouabain; D008715:Methionine; D000254:Sodium-Potassium-Exchanging ATPase; D002945:Cisplatin
keywords: Adverse effects; Binding sites; Cisplatin; Na(+)/K(+)-ATPase; Sodium pump; X-ray crystallography