Expression, Purification, and Metal Binding Properties of the N-terminal Domain from the Wilson Disease Putative Copper-transporting ATPase (ATP7B)*

The putative copper binding domain from the copper-transporting ATPase implicated in Wilson disease (ATP7B) has been expressed and purified as a fusion to glutathioneS-transferase. Immobilized metal ion affinity chromatography revealed that the fusion protein is able to bind to columns charged with different transition metals with varying affinities as follows: Cu(II)≫Zn(II)>Ni(II)>Co(II). The fusion protein did not bind to columns charged with Fe(II) or Fe(III).65Zinc(II) blotting analysis showed that the domain is able to bind Zn(II) over a range of pH values from 6.5 to 9.0. Competition65Zn(II) blotting showed that Cd(II), Hg(II), Au(III), and Fe(III) can successfully compete with Zn(II), at comparable concentrations, for binding to the domain. In contrast, the domain had little or no affinity for Ca(II), Mg(II), Mn(II), and Ni(II) relative to copper. Neutron activation analysis of the copper bound to the domain showed a copper:protein ratio of 6.5–7.3:1. Both Cu(II) and Cu(I) were found to have a higher affinity for the domain relative to Zn(II). In addition, a sharp, reproducible transition was only observed in competition experiments with copper, which may suggest that copper binding has some degree of cooperativity.

• Publication year

  1997

• Venue

  Journal of Biological Chemistry

• Publication date

  1997-12-26

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/jbc.272.52.33279PMID 9407118
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• N-terminal Domains of Human Copper-transporting Adenosine Triphosphatases (the Wilson’s and Menkes Disease Proteins) Bind Copper Selectively in Vivo and in Vitro with Stoichiometry of One Copper Per Metal-binding Repeat*

  1997cited by this paper
• Copper transport and its alterations in Menkes and Wilson diseases.

  1997cited by this paper
• Biochemical characterization and intracellular localization of the Menkes disease protein.

  1996cited by this paper
• In Vivo and in Vitro Iron-replaced Zinc Finger Generates Free Radicals and Causes DNA Damage (*)

  1996cited by this paper
• Characterization of calf liver Cu,Zn-metallothionein: naturally variable Cu and Zn stoichiometries.

  1996cited by this paper
• Ligand‐regulated transport of the Menkes copper P‐type ATPase efflux pump from the Golgi apparatus to the plasma membrane: a novel mechanism of regulated trafficking.

  1996cited by this paper
• An unusual metal‐binding cluster found exclusively in the avian breast muscle troponin T of Galliformes and Craciformes

  1994cited by this paper
• Induction of the putative copper ATPases, CopA and CopB, of Enterococcus hirae by Ag+ and Cu2+, and Ag+ extrusion by CopB.

  1994cited by this paper
• P-type ATPase from the cyanobacterium Synechococcus 7942 related to the human Menkes and Wilson disease gene products.

  1994cited by this paper
• The Wilson disease gene is a copper transporting ATPase with homology to the Menkes disease gene

  1993cited by this paper
• The Wilson disease gene is a putative copper transporting P–type ATPase similar to the Menkes gene

  1993cited by this paper
• Isolation of a partial candidate gene for Menkes disease by positional cloning

  1993cited by this paper
• Isolation of a candidate gene for Menkes disease and evidence that it encodes a copper–transporting ATPase

  1993cited by this paper
• Isolation of a candidate gene for Menkes disease that encodes a potential heavy metal binding protein

  1993cited by this paper
• Transition metals in control of gene expression.

  1993cited by this paper
• Polypeptides containing highly conserved regions of transcription initiation factor sigma 70 exhibit specificity of binding to promoter DNA.

  1992cited by this paper
• Bacterial resistance ATPases: primary pumps for exporting toxic cations and anions.

  1989cited by this paper
• The Metabolic basis of inherited disease

  1989cited by this paper
• Characterization of a zinc blotting technique: evidence that a retroviral gag protein binds zinc.

  1988cited by this paper
• Stability of enzymes.

  1985cited by this paper
• Protein purification: Principles and practice

  1984cited by this paper

• Tackling cuproptosis: from metabolic rewiring to therapeutic exploitation in cancer

  2026cites this paper
• Cuproptosis in cancer: from molecular mechanisms to therapeutic intervention.

  2025cites this paper
• Copper in cancer: friend or foe? Metabolism, dysregulation, and therapeutic opportunities

  2025cites this paper
• A metabolic perspective on cuproptosis.

  2025cites this paper
• Structure and mechanism of the human copper transporting ATPases: Fitting the pieces into a moving puzzle.

  2024cites this paper
• Targeting cuproplasia and cuproptosis in cancer

  2024cites this paper
• Genome-wide association studies and functional annotation of pre-weaning calf mortality and reproductive traits in Nellore cattle from experimental selection lines

  2024cites this paper
• Copper Metabolism and Cuproptosis: Molecular Mechanisms and Therapeutic Perspectives in Neurodegenerative Diseases

  2024cites this paper
• Bioaccumulation of mercury and transcriptional responses in tusk (Brosme brosme), a deep-water fish from a Norwegian fjord.

  2021cites this paper
• ScholarWorks at WMU ScholarWorks at WMU Copper Exchange Studies between the First and Fourth N- Copper Exchange Studies between the First and Fourth N-Terminal Metal-Binding Domains of Wilson Disease ATPase Terminal Metal-Binding Domains of Wilson Disease ATPase

  2021cites this paper
• Copper Dyshomeostasis in Neurodegenerative Diseases—Therapeutic Implications

  2020cites this paper
• Structural Role of the First Four Transmembrane Helices in ZntA, a P1B-Type ATPase from Escherichia coli.

  2020cites this paper
• Benchmarking Computational Methods for Estimating the Pathogenicity of Wilson’s Disease Mutations

  2019cites this paper
• Long-Term Correction of Copper Metabolism in Wilson's Disease Mice with AAV8 Vector Delivering Truncated ATP7B

  2019cites this paper
• Biochemical and Cellular Properties of ATP7B Variants

  2019cites this paper
• Computing the Pathogenicity of Wilson's Disease ATP7B Mutations: Implications for Disease Prevalence

  2019cites this paper
• Wilson ’ s Disease Protein

  2019cites this paper
• Molecular Architecture of the Copper-Transporting ATPase ATP7B

  2019cites this paper
• Copper-dependent ATP7B upregulation drives the resistance of TMEM16A-overexpressing head-and-neck cancer models to platinum toxicity.

  2019cites this paper
• A glimpse into the regulation of the Wilson disease protein, ATP7B, sheds light on the complexity of mammalian apical trafficking pathways.

  2018cites this paper
• Cellular copper homeostasis: current concepts on its interplay with glutathione homeostasis and its implication in physiology and human diseases.

  2017cites this paper
• Investigating the role of cua-1 in maintaining copper homeostasis within a long-lived, insulin-signalling, C. elegans mutants

  2017cites this paper
• 2 Biochemistry of the Wilson’s Disease Protein

  2017cites this paper
• The six metal binding domains in human copper transporter, ATP7B: molecular biophysics and disease-causing mutations

  2017cites this paper
• Molecular features of copper binding proteins involved in copper homeostasis

  2017cites this paper
• Broadening the implications of gene discovery

  2016cites this paper
• Interactions between Metal-binding Domains Modulate Intracellular Targeting of Cu(I)-ATPase ATP7B, as Revealed by Nanobody Binding*

  2014cites this paper
• Biochemical characterization of P-type copper ATPases

  2014cites this paper
• Visualizing Hepatic Copper Release in Long–Evans Cinnamon Rats Using Single-Photon Emission Computed Tomography

  2013cites this paper
• Copper Transporting ATPases in Mammalian Cells

  2013cites this paper
• Copper chaperone Atox1 interacts with the metal-binding domain of Wilson's disease protein in cisplatin detoxification.

  2013cites this paper
• Structural and Functional Studies of ATP7B, the Copper(I)-transporting P-type ATPase Implicated in Wilson Disease

  2012cites this paper
• Metal‐Related Diseases of Genetic Origin

  2011cites this paper
• Mechanism of tumor resistance to cisplatin mediated by the copper transporter ATP7B.

  2011cites this paper
• The puzzle posed by COMMD1, a newly discovered protein binding Cu(II).

  2011cites this paper
• Wilson's disease.

  2011cites this paper
• Association between the c. 2495 A>G ATP7B Polymorphism and Sporadic Alzheimer's Disease

  2011cites this paper
• Copper Transporters and Chaperones

  2011cites this paper
• Copper-transfer mechanism from the human chaperone Atox1 to a metal-binding domain of Wilson disease protein.

  2010cites this paper
• NMR characterization of copper-binding domains 4-6 of ATP7B .

  2010cites this paper
• A Combined Zinc/Cadmium Sensor and Zinc/Cadmium Export Regulator in a Heavy Metal Pump*

  2010cites this paper
• Interdomain interactions modulate collective dynamics of the metal-binding domains in the Wilson disease protein.

  2010cites this paper
• Characterization of the N-Terminal Domains and Disease-Causing Mutations of the Human Wilson Protein

  2010cites this paper
• Structural organization of human Cu-transporting ATPases: learning from building blocks

  2009cites this paper
• Functional Interactions of Cu-ATPase ATP7B with Cisplatin and the Role of ATP7B in the Resistance of Cells to the Drug*

  2009cites this paper
• Therapeutic Targeting of ATP7B in Ovarian Carcinoma

  2009cites this paper
• Functional analysis of the heavy metal binding domains of the Zn/Cd-transporting ATPase, HMA2, in Arabidopsis thaliana.

  2009cites this paper
• Cadmium-mediated rescue from ER-associated degradation induces expression of its exporter

  2009influential citation
• The ligand dependent interactions between cytoplasmic domains in Cu+ transporter, Archaeoglobus fulgidus

  2009cites this paper
• Conformational dynamics of metal-binding domains in Wilson disease protein: molecular insights into selective copper transfer.

  2009cites this paper
• Copper as a biological yin-yang element: Structural dynamics, protein -protein interactions and transfer mechanisms of copper transport proteins

  2009cites this paper
• Metal binding domains 3 and 4 of the Wilson disease protein: solution structure and interaction with the copper(I) chaperone HAH1.

  2008cites this paper
• Characterization of the role of copper transporting ATPase, ATP7B (Wilsons disease protein), in copper homeostasis.

  2008cites this paper
• Evidence for direct interactions between the mercuric ion transporter (MerT) and mercuric reductase (MerA) from the Tn501mer operon

  2008cites this paper
• Liver Ischemia and Ischemia–Reperfusion Induces and Trafficks the Multi-specific Metal Transporter Atp7b to Bile Duct Canaliculi: Possible Preferential Transport of Iron into Bile

  2008cites this paper
• Molecular characterization of the copper transport system in Staphylococcus aureus.

  2007cites this paper
• The structure and function of heavy metal transport P1B-ATPases

  2007influential citation
• Conservative and nonconservative mutations of the transmembrane CPC motif in ZntA: effect on metal selectivity and activity.

  2007cites this paper
• HMA2. A Transmembrane Zn 2+ Transporting ATPase from Arabidopsis thaliana

  2007cites this paper
• Immobilized Metal‐Ion Affinity Chromatography: Status and Trends

  2007cites this paper
• Function and regulation of human copper-transporting ATPases.

  2007cites this paper
• Molecular pathogenesis of Wilson and Menkes disease: correlation of mutations with molecular defects and disease phenotypes

  2007cites this paper
• Cu(I) Binding and Transfer by the N Terminus of the Wilson Disease Protein*

  2007cites this paper
• Searching for Genes Involved in Metal Tolerance, Uptake, and Transport

  2007cites this paper
• Characterization and copper binding properties of human COMMD1 (MURR1).

  2007cites this paper
• Purification and membrane reconstitution of catalytically active Menkes copper-transporting P-type ATPase (MNK; ATP7A).

  2007cites this paper
• Efficacy of Chemotherapeutic Agents Under Hypoxic Conditions in Pulmonary Adenocarcinoma Multidrug Resistant Cell Line

  2007cites this paper
• Efficacy of chemotherapeutic agents under hypoxic conditions in pulmonary adenocarcinoma multidrug resistant cell line.

  2007cites this paper
• Novel Zn2+ coordination by the regulatory N-terminus metal binding domain of Arabidopsis thaliana Zn(2+)-ATPase HMA2.

  2007cites this paper
• Regulation of the Cellular Pharmacology and Cytotoxicity of Cisplatin by Copper Transporters

  2006cites this paper
• Role of metal-binding domains of the copper pump from Archaeoglobus fulgidus.

  2006cites this paper
• Proteomics of metal transport and metal-associated diseases.

  2006cites this paper
• Copper-dependent Interaction of Dynactin Subunit p62 with the N Terminus of ATP7B but Not ATP7A*

  2006cites this paper
• A Novel Regulatory Metal Binding Domain Is Present in the C Terminus of Arabidopsis Zn2+-ATPase HMA2*

  2006cites this paper
• Structure of human Wilson protein domains 5 and 6 and their interplay with domain 4 and the copper chaperone HAH1 in copper uptake.

  2006cites this paper
• Metal-binding affinity of the transmembrane site in ZntA: implications for metal selectivity.

  2006cites this paper
• Kinetic analysis of metal binding to the amino-terminal domain of ZntA by monitoring metal-thiolate charge-transfer complexes.

  2005cites this paper
• Contribution of Va24Vb11 natural killer T cells in Wilsonian hepatitis.

  2005cites this paper
• Metal-binding characteristics of the amino-terminal domain of ZntA: binding of lead is different compared to cadmium and zinc.

  2005cites this paper
• Copper Exchange Studies between the First and Fourth N-Terminal Metal-Binding Domains of Wilson Disease ATPase

  2005cites this paper
• Confocal Microscopic Analysis of the Interaction between Cisplatin and the Copper Transporter ATP7B in Human Ovarian Carcinoma Cells

  2004cites this paper
• Intracellular trafficking of the human Wilson protein: the role of the six N-terminal metal-binding sites.

  2004cites this paper
• Binding of Copper(I) by the Wilson Disease Protein and Its Copper Chaperone*

  2004cites this paper
• Identification of the "missing domain" of the rat copper-transporting ATPase, atp7b: insight into the structural and metal binding characteristics of its N-terminal copper-binding domain.

  2004cites this paper
• The N-terminal Metal-binding Site 2 of the Wilson's Disease Protein Plays a Key Role in the Transfer of Copper from Atox1*

  2004cites this paper
• Copper transfer to the N-terminal domain of the Wilson disease protein (ATP7B): X-ray absorption spectroscopy of reconstituted and chaperone-loaded metal binding domains and their interaction with exogenous ligands.

  2004cites this paper
• Influence of transition and heavy metal ions on ATP-ases activity in rat synaptic plasma membranes

  2004cites this paper
• Kinetic Analysis of the Interaction of the Copper Chaperone Atox1 with the Metal Binding Sites of the Menkes Protein*

  2003cites this paper
• Current and prospective applications of metal ion-protein binding.

  2003cites this paper
• Identification of Ion-Selectivity Determinants in Heavy-Metal Transport P1B-type ATPases

  2003cites this paper
• Cellular copper transport and metabolism.

  2003cites this paper
• Function and Regulation of the Mammalian Copper-transporting ATPases: Insights from Biochemical and Cell Biological Approaches

  2003cites this paper
• X‐ray crystal structure of CutA from Thermotoga maritima at 1.4 Å resolution

  2003cites this paper
• Structural Basis for the Function of the N-terminal Domain of the ATPase CopA from Bacillus subtilis*

  2003cites this paper
• Functional roles of metal binding domains of the Archaeoglobus fulgidus Cu(+)-ATPase CopA.

  2003cites this paper
• A novel Nα-acetyl alanine aminopeptidase from Allomyces arbuscula

  2002cites this paper
• Microbial Resistance Mechanisms for Heavy Metals and Metalloids

  2002cites this paper
• Biochemical Characterization of CopA, the Escherichia coli Cu(I)-translocating P-type ATPase*

  2002cites this paper
• A novel N(alpha)-acetyl alanine aminopeptidase from Allomyces arbuscula.

  2002cites this paper
• Characterization of a Thermophilic P-type Ag+/Cu+-ATPase from the ExtremophileArchaeoglobus fulgidus*

  2002cites this paper