Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution

Rosenzweig, Amy C.; Huffman, David L.; Hou, Melody Y.; Wernimont, A.K.; Pufahl, Robert A.; O’Halloran, Thomas V.

doi:10.1016/s0969-2126(99)80082-3

Cited by 233 publications

(292 citation statements)

References 47 publications

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“…Sequence alignments and homology models on the six metal-binding domains of WLNP demonstrate that the Ϸ70 aa domains of N-WLNP are likely to be folded very similarly into ferredoxin-like units (16). This same fold is also found in both NMR and x-ray structures of the yeast Atx1 and human HAH1 metallochaperones (17)(18)(19)(20), the soluble cytosolic proteins that deliver copper to the copper-transporting P-type ATPases (21).…”

mentioning

confidence: 67%

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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mentioning

confidence: 67%

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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“…This residue, which is essential for function (58), has been proposed to modulate copper transfer (50,52). In the CuCox17 structure, conserved residue Lys 30 is proximal to the copper-binding site and could counterbalance the negative charge because of the presence of three cysteines.…”

Section: Resultsmentioning

confidence: 99%

Yeast Cox17 Solution Structure and Copper(I) Binding

Abajian

Yatsunyk

Ramirez

et al. 2004

Journal of Biological Chemistry

100

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“…Consideration of the electrostatic properties of each domain furnishes part of the answer. Interactions between Atox1 or Atx1 and their target domains involve complementary electrostatic surfaces (15,16,37,48). In particular, positively charged residues on the chaperone surface interact with negatively charged residues on the target domain.…”

Section: Discussionmentioning

confidence: 99%

“…The Atox1 metallochaperone and its homologs, both eukaryotic and prokaryotic, contain a single CXXC motif (14). Crystal and solution structures of Atox1 (15), yeast Atx1 (16,17), bacterial CopZ (18,19), and single domains of MNK (20) and Ccc2 (21) reveal a conserved ␤␣␤␤␣␤-fold with the cysteines from the CXXC motif coordinating metal ions on a surface-exposed loop. Notably, the CXXC motifs from two Atox1 molecules coordinate a single Cu(I) ion in the x-ray structure (15).…”

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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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Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution

Cited by 233 publications

References 47 publications

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

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

Yeast Cox17 Solution Structure and Copper(I) Binding

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

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