Chapter Mechanism of Human Copper Transporter Wilson’s Disease Protein

2016 ◽  
pp. 159-180
Keyword(s):  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Copper Transporter ◽  
Disease Protein

The Wilson's disease protein expressed in Sf9 cells is phosphorylated

2002 ◽  
Vol 30 (4) ◽  
pp. 739-741 ◽  
Author(s):  
S. M. Vanderwerf ◽  
S. Lutsenko
Keyword(s):  
Mammalian Cells ◽  
Wilson’S Disease ◽  
Insect Cells ◽  
Physiological Role ◽  
Wilson's Disease ◽  
Copper Transporter ◽  
Mammalian Cell Lines ◽  
Disease Protein ◽  
Baculovirus System

The Wilson's disease protein (WNDP), a copper transporter, is a crucial mediator of copper homoeostasis in mammalian cells. We recently found that changes in copper concentration regulate the phosphorylation level of WNDP. WNDP phosphorylation was observed in several mammalian cell lines, suggesting that a common phosphorylation pathway exists in these cells. Here we demonstrate that WNDP expressed in Sf9 insect cells is also phosphorylated, as evidenced by metabolic labelling of these cells with [32P]Pi. Because the baculovirus system allows us to generate large amounts of protein, we are using this expression method to isolate WNDP and map the sites of WNDP phosphorylation. The identification of phosphorylation sites is the first step towards understanding the physiological role of WNDP phosphorylation.

Copper chaperone Atox1 interacts with the metal-binding domain of Wilson's disease protein in cisplatin detoxification

2013 ◽  
Vol 454 (1) ◽  
pp. 147-156 ◽  
Author(s):  
Nataliya V. Dolgova ◽  
Sergiy Nokhrin ◽  
Corey H. Yu ◽  
Graham N. George ◽  
Oleg Y. Dmitriev
Keyword(s):  
Metal Binding ◽  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Binding Domain ◽  
Binding Motif ◽  
Copper Chaperone ◽  
Copper Binding ◽  
Copper Transporters ◽  
Metal Binding Domain ◽  
Disease Protein

Human copper transporters ATP7B (Wilson's disease protein) and ATP7A (Menkes' disease protein) have been implicated in tumour resistance to cisplatin, a widely used anticancer drug. Cisplatin binds to the copper-binding sites in the N-terminal domain of ATP7B, and this binding may be an essential step of cisplatin detoxification involving copper ATPases. In the present study, we demonstrate that cisplatin and a related platinum drug carboplatin produce the same adduct following reaction with MBD2 [metal-binding domain (repeat) 2], where platinum is bound to the side chains of the cysteine residues in the CxxC copper-binding motif. This suggests the same mechanism for detoxification of both drugs by ATP7B. Platinum can also be transferred to MBD2 from copper chaperone Atox1, which was shown previously to bind cisplatin. Binding of the free cisplatin and reaction with the cisplatin-loaded Atox1 produce the same protein-bound platinum intermediate. Transfer of platinum along the copper-transport pathways in the cell may serve as a mechanism of drug delivery to its target in the cell nucleus, and explain tumour-cell resistance to cisplatin associated with the overexpression of copper transporters ATP7B and ATP7A.

scholarly journals Functional Properties of the Copper-transporting ATPase ATP7B (The Wilson's Disease Protein) Expressed in Insect Cells

2001 ◽  
Vol 277 (2) ◽  
pp. 976-983 ◽  
Author(s):  
Ruslan Tsivkovskii ◽  
John F. Eisses ◽  
Jack H. Kaplan ◽  
Svetlana Lutsenko
Keyword(s):  
Functional Properties ◽  
Wilson’S Disease ◽  
Insect Cells ◽  
Wilson's Disease ◽  
Disease Protein

scholarly journals Molecular modelling of the nucleotide-binding domain of Wilson's disease protein: location of the ATP-binding site, domain dynamics and potential effects of the major disease mutations

2004 ◽  
Vol 382 (1) ◽  
pp. 293-305 ◽  
Author(s):  
Roman G. EFREMOV ◽  
Yuri A. KOSINSKY ◽  
Dmitry E. NOLDE ◽  
Ruslan TSIVKOVSKII ◽  
Alexander S. ARSENIEV ◽  
...  
Keyword(s):  
Molecular Dynamics ◽  
Molecular Dynamics Simulations ◽  
Molecular Modelling ◽  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Binding Domain ◽  
Atp Binding ◽  
Disease Mutations ◽  
Disease Protein ◽  
Dynamics Simulations

WNDP (Wilson's disease protein) is a copper-transporting ATPase that plays an essential role in human physiology. Mutations in WNDP result in copper accumulation in tissues and cause a severe hepato-neurological disorder known as Wilson's disease. Several mutations were surmised to affect the nucleotide binding and hydrolysis by WNDP; however, how the nucleotides bind to normal and mutated WNDP remains unknown. To aid such studies, we performed the molecular modelling of the spatial structure and dynamics of the ATP-binding domain of WNDP and its interactions with ATP. The three-dimensional models of this domain in two conformations were built using the X-ray structures of the Ca2+-ATPase in the E1 and E2 states. To study the functional aspects of the models, they were subjected to long-term molecular dynamics simulations in an explicit solvent; similar calculations were performed for the ATP-binding domain of Ca2+-ATPase. In both cases, we found large-scale motions that lead to significant changes of distances between several functionally important residues. The ATP docking revealed two possible modes of ATP binding: via adenosine buried in the cleft near residues H1069, R1151 and D1164, and via phosphate moiety ‘anchored’ by H-bonds with residues in the vicinity of catalytic D1027. Furthermore, interaction of ATP with both sites occurs if they are spatially close to each other. This may be achieved after relative domain motions of the ‘closure’ type observed in molecular dynamics simulations. The results provide a framework for analysis of disease mutations and for future mutagenesis studies.

Biochemistry of the Wilson's Disease Protein

2003 ◽  
pp. 035-051
Author(s):  
Svetlana Lutsenko ◽  
Ruslan Tsivkovskii ◽  
Matthew J. Cooper ◽  
Brian C. MacArthur ◽  
Hans-Peter Bächinger
Keyword(s):  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Disease Protein

Localization of the Wilson's disease protein in human liver

1999 ◽  
Vol 117 (6) ◽  
pp. 1380-1385 ◽  
Author(s):  
Mark Schaefer ◽  
Han Roelofsen ◽  
Henk Wolters ◽  
Walter J. Hofmann ◽  
Michael Müller ◽  
...  
Keyword(s):  
Human Liver ◽  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Disease Protein

Hepatocyte-specific localization and copper-dependent trafficking of the Wilson’s disease protein in the liver

1999 ◽  
Vol 276 (3) ◽  
pp. G639-G646 ◽  
Author(s):  
Mark Schaefer ◽  
Robin G. Hopkins ◽  
Mark L. Failla ◽  
Jonathan D. Gitlin
Keyword(s):  
Wilson’S Disease ◽  
Neuronal Degeneration ◽  
Copper Metabolism ◽  
Wilson's Disease ◽  
Polyclonal Antiserum ◽  
Copper Excretion ◽  
Hepatic Expression ◽  
Hepatic Copper ◽  
Disease Protein

Wilson’s disease is an inherited disorder of copper metabolism characterized by hepatic cirrhosis and neuronal degeneration. In this current study, a polyclonal antiserum specific for the Wilson’s disease ATPase was used to examine the hepatic expression of this protein. Immunoblot analysis of lysates from human and rat liver detected a single 165-kDa protein, which by immunofluorescence was present only in hepatocytes and localized predominantly to the trans-Golgi network and exclusively in this compartment under low hepatic copper concentrations. Although hepatic copper concentration had no effect on the steady-state levels of the Wilson’s disease protein, copper administration in vivo resulted in redistribution of this protein to a cytoplasmic vesicular compartment localized toward the hepatocyte canalicular membrane. The relative abundance of the Wilson’s disease protein in the liver was found to be greatest in the fetus before the onset of biliary copper excretion. Taken together, these studies reveal a novel posttranslational mechanism of copper homeostasis in vivo consistent with the proposed function of the Wilson’s disease protein in holoceruloplasmin biosynthesis and biliary copper excretion and of relevance to the broad clinical heterogeneity observed in this disease.

Sign in / Sign up

Export Citation Format

Share Document