Analysis of functional domains of Wilson disease protein (ATP7B) in <i>Saccharomyces cerevisiae</i>

Iida, Masatake; Terada, Kunihiko; Sambongi, Yoshihiro; Wakabayashi, Tokumitsu; Miura, Naoyuki; Koyama, Kenji; Futai, Masamitsu; Sugiyama, Toshihiro

doi:10.1016/s0014-5793(98)00546-8

Cited by 96 publications

(91 citation statements)

References 26 publications

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“…Currently, the most frequently used system for analysis of WNDP and its disease-related mutants utilizes a yeast strain, in which the endogenous copper-transporting ATPase CCC2 has been deleted (30). Deletion of CCC2 disrupts copper delivery to the secretory compartment and renders the copper-dependent ferroxidase Fet3 inactive, while expression of WNDP restores the Fet3 activity to some extent (18,31). This elegant assay permits rapid screening of mutants (32), however, the indirect nature of the assay prevents characterization of the molecular mechanism of WNDP.…”

Section: Characterization Of the Wndp Expression In Insect Cells-mentioning

confidence: 99%

“…The observed cooperativity could be either due to interactions between different intramembrane copper-binding sites or due to cross-talk between the intramembrane copper-binding site and the site(s) at the N-terminal portion of WNDP. The latter explanation is very appealing since previous studies demonstrated that the deletion/mutations of the 5th and/or 6th Nterminal metal-binding sites inhibited transport activity of WNDP (31,47).…”

Section: Functional Expression Of Copper-atpase Atp7bmentioning

confidence: 99%

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

Tsivkovskii¹,

Eisses²,

Kaplan

et al. 2002

Journal of Biological Chemistry

100

133

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Section: Characterization Of the Wndp Expression In Insect Cells-mentioning

confidence: 99%

Section: Functional Expression Of Copper-atpase Atp7bmentioning

confidence: 99%

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

Tsivkovskii¹,

Eisses²,

Kaplan

et al. 2002

Journal of Biological Chemistry

100

133

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“…Mutational studies performed to address the role of the N-WLNP have found that, generally, the domains closest to the membrane seem to be the most important both for the ultimate incorporation of copper into Fet3 in yeast complementation assays and in the copper-dependent translocation of WLNP from the vesicular to the cytosolic membrane (23)(24)(25). However, the mechanism by which these domains acquire copper and then transport it across the vesicular membrane is unknown.…”

mentioning

confidence: 99%

Structure of human Wilson protein domains 5 and 6 and their interplay with domain 4 and the copper chaperone HAH1 in copper uptake

Achila

Banci

Bertini

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

146

252

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Human Wilson protein is a copper-transporting ATPase located in the secretory pathway possessing six N-terminal metal-binding domains. Here we focus on the function of the metal-binding domains closest to the vesicular portion of the copper pump, i.e., domain 4 (WLN4), and a construct of domains 5 and 6 (WLN5-6). For comparison purposes, some experiments were also performed with domain 2 (WLN2). The solution structure of apoWLN5-6 consists of two ferredoxin folds connected by a short linker, and 15 N relaxation rate measurements show that it behaves as a unit in solution. An NMR titration of apoWLN5-6 with the metallochaperone Cu(I)HAH1 reveals no complex formation and no copper exchange between the two proteins, whereas titration of Cu(I)HAH1 with WLN4 shows the formation of an adduct that is in fast exchange on the NMR time scale with the isolated protein species as confirmed by 15 N relaxation data. A similar interaction is also observed between Cu(I)HAH1 and WLN2; however, the relative amount of the adduct in the protein mixture is lower. An NMR titration of apoWLN5-6 with Cu(I)WLN4 shows copper transfer, first to WLN6 then to WLN5, without the formation of an adduct. Therefore, we suggest that WLN4 and WLN2 are two acceptors of Cu(I) from HAH1, which then somehow route copper to WLN5-6, before the ATP-driven transport of copper across the vesicular membrane.ATPase function ͉ copper transport ͉ metal-binding domain ͉ metallochaperone ͉ Wilson disease protein

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“…Numerous genetic and biochemical studies suggest that the domains are not functionally equivalent. For WND, yeast complementation assays indicate that the sixth domain alone is sufficient for copper loading of Fet3, the yeast homolog of ceruloplasmin (26,27). Furthermore, the second or third metal-binding domain cannot substitute for the sixth domain, supporting distinct functions for the individual domains (27).…”

mentioning

confidence: 99%

Binding of Copper(I) by the Wilson Disease Protein and Its Copper Chaperone

Wernimont¹,

Yatsunyk

Rosenzweig

2004

Journal of Biological Chemistry

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The Wilson disease protein (WND) is a transport ATPase involved in copper delivery to the secretory pathway. Mutations in WND and its homolog, the Menkes protein, lead to genetic disorders of copper metabolism. The WND and Menkes proteins are distinguished from other P-type ATPases by the presence of six soluble N-terminal metal-binding domains containing a conserved CXXC metal-binding motif. The exact roles of these domains are not well established, but possible functions include exchanging copper with the metallochaperone Atox1 and mediating copper-responsive cellular relocalization. Although all six domains can bind copper, genetic and biochemical studies indicate that the domains are not functionally equivalent. One way the domains could be tuned to perform different functions is by having different affinities for Cu(I). We have used isothermal titration calorimetry to measure the association constant (K a ) and stoichiometry (n) values of Cu(I) binding to the WND metal-binding domains and to their metallochaperone Atox1. The association constants for both the chaperone and target domains are ϳ10 5 to 10 6 M ؊1 , suggesting that the handling of copper by Atox1 and copper transfer between Atox1 and WND are under kinetic rather than thermodynamic control. Although some differences in both n and K a values are observed for variant proteins containing less than the full complement of six metal-binding domains, the data for domains 1-6 were best fitted with a single site model. Thus, the individual functions of the six WND metal-binding domains are not conferred by different Cu(I) affinities but instead by fold and electrostatic surface properties.

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Analysis of functional domains of Wilson disease protein (ATP7B) in Saccharomyces cerevisiae

Cited by 96 publications

References 26 publications

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

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

Structure of human Wilson protein domains 5 and 6 and their interplay with domain 4 and the copper chaperone HAH1 in copper uptake

Binding of Copper(I) by the Wilson Disease Protein and Its Copper Chaperone

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