Copper relay path through the N-terminus of Wilson disease protein, ATP7B

Shanmugavel, Kumaravel Ponnandai; Wittung-Stafshede, Pernilla

doi:10.1039/c9mt00147f

Cited by 21 publications

(32 citation statements)

References 42 publications

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“…This study shows that Atox1 interacts with MBD1-3, where it is thought that Cu(I) is initially loaded to MBD2, then Cu(I) is transferred from MBD3 to MBD6. Another scenario was suggested whereby Atox1 interacts with MBD4, and then Cu(I) is transferred to MBD6 [16]. The EPR performed here supports the conclusion that MBD1-3 and MBD4-6 act as independent units.…”

Section: Discussionsupporting

confidence: 85%

“…However, experimental and computational research suggests that Atox1 can form a stable adduct with MBD1, MBD2 and MBD4, which are more solvent exposed; however, not with MBD3 and MBD5-6 [12,[21][22][23]. NMR experiments, yeast cell experiments and MD simulations suggest that MBD1-4 are much more dynamic than MBD5-6 [11][12][13]15,16]. These studies also proposed that MBD4 has little interaction with MBD1-3 and MBD5-6.…”

Section: Discussionmentioning

confidence: 99%

“…Molecular dynamics (MD) simulation suggests that MBD1-3 and MBD5-6 act as two independent units, whereas MBD4 has little interaction with each of them [ 15 ]. Yeast cell experiments showed that MBD1-3 acts as a regulatory unit, whereas MBD5-6 is essential for ensuring proper copper transfer [ 16 ]. The importance of MBD5-6 in copper transfer is also linked to Wilson’s disease, wherein most of the mutations in the N-terminus domain of ATP7B were reported in this unit [ 17 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

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An EPR Study on the Interaction between the Cu(I) Metal Binding Domains of ATP7B and the Atox1 Metallochaperone

Zaccak

Qasem

Gevorkyan‐Airapetov

et al. 2020

IJMS

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Copper’s essentiality and toxicity mean it requires a sophisticated regulation system for its acquisition, cellular distribution and excretion, which until now has remained elusive. Herein, we applied continuous wave (CW) and pulsed electron paramagnetic resonance (EPR) spectroscopy in solution to resolve the copper trafficking mechanism in humans, by considering the route travelled by Cu(I) from the metallochaperone Atox1 to the metal binding domains of ATP7B. Our study revealed that Cu(I) is most likely mediated by the binding of the Atox1 monomer to metal binding domain 1 (MBD1) and MBD4 of ATP7B in the final part of its extraction pathway, while the other MBDs mediate this interaction and participate in copper transfer between the various MBDs to the ATP7B membrane domain. This research also proposes that MBD1-3 and MBD4-6 act as two independent units.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An EPR Study on the Interaction between the Cu(I) Metal Binding Domains of ATP7B and the Atox1 Metallochaperone

Zaccak

Qasem

Gevorkyan‐Airapetov

et al. 2020

IJMS

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“…Mammalian copper ATPases harbours six such motifs on the amino terminus. This motif sequesters copper and facilitates its transport across the membrane to the lumen [30]. Using homology modelling based on structures of the amino-terminal HM domains of metal ATPases available in the Protein database (PDB), we determine if the novel ATPase that we predicted in the Leishmania genome belongs to the family of heavy metal transporting ATPases.…”

Section: Resultsmentioning

confidence: 99%

The novel leishmanial Copper P-type ATPase plays a vital role in intracellular parasite survival

Paul

Banerjee

Sen

et al. 2021

Preprint

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Copper is essential for all life forms; however in excess it is extremely toxic. Toxic properties of copper are utilized by hosts against various pathogenic invasions. Leishmania, in its both free-living and intracellular forms was found to exhibit appreciable tolerance towards copper-stress. To determine the mechanism of copper-stress evasion employed by Leishmania we identified and characterized the hitherto unknown Copper-ATPase in Leishmania major and determined its role in parasite’s survival in host macrophage cells. L. major Cu-ATPase, LmATP7, exhibits high homology with its orthologues at multiple conserved motifs. In promastigotes, LmATP7 localized to the plasma membrane with a fraction in intracellular puncta. Upon copper treatment, LmATP7 expression increases few folds. LmATP7 is capable of complementing copper transport in Cu-ATPase-Δ yeast strain. Promastigotes overexpressing LmATP7 exhibits higher survival upon copper stress indicating efficacious copper export compared to wild type parasites. We explored macrophage-Leishmanial interaction with respect to copper stress subjected by the host upon parasite and the parasite’s reciprocating response thereon to evade the stress. The Copper-P-type-ATPases ATP7A/7B serves as major copper exporter in eukaryotes that maintain cellular copper levels. We found that Leishmania infection, triggers ATP7A upregulation in macrophages. Additionally, as part of host response, ATP7A traffics from trans-Golgi network and transports copper to the endosomal and endolysosomal compartments harbouring the Leishmania amastigotes. Finally, we show LmATP7 overexpression in this parasite, increased amastigote survivability within infected macrophages, establishing its role in combating host-induced copper stress.

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“…Probing ATP7B-mediated Cu-transport in a yeast assay If an ATP7B mutation causes WD, the obvious speculation would be that the mutation hampers Cu transport by the mutated ATP7B protein. Therefore, to investigate Cu transport ability by the four mutated ATP7B proteins, we turned to a yeast assay that has been used by many to assess ATP7B dysfunction (Forbes et al 1999;Ponnandai Shanmugavel et al 2017) and that was improved by us recently to also include Cu delivery from human Atox1 (Ponnandai Shanmugavel et al 2017;Shanmugavel and Wittung-Stafshede 2019). In this set up, the Dccc2Datx1 double knockout yeast strain supplemented with the human ATOX1 gene acted as the background strain to which constructs with mutated ATP7B were introduced on high copy plasmids for the constitutive expression of mutant ATP7B proteins.…”

Section: Resultsmentioning

confidence: 99%

Wilson disease missense mutations in ATP7B affect metal-binding domain structural dynamics

Shanmugavel

Kumar

et al. 2019

Biometals

Self Cite

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Wilson disease (WD) is caused by mutations in the gene for ATP7B, a copper transport protein that regulates copper levels in cells. A large number of missense mutations have been reported to cause WD but genotype-phenotype correlations are not yet established. Since genetic screening for WD may become reality in the future, it is important to know how individual mutations affect ATP7B function, with the ultimate goal to predict pathophysiology of the disease. To begin to assess mechanisms of dysfunction, we investigated four proposed WD-causing missense mutations in metal-binding domains 5 and 6 of ATP7B. Three of the four variants showed reduced ATP7B copper transport ability in a traditional yeast assay. To probe mutation-induced structural dynamic effects at the atomic level, molecular dynamics simulations (1.5 ls simulation time for each variant) were employed. Upon comparing individual metal-binding domains with and without mutations, we identified distinct differences in structural dynamics via root-mean square fluctuation and secondary structure content analyses. Most mutations introduced distant effects resulting in increased dynamics in the copper-binding loop. Taken together, mutation-induced long-range alterations in structural dynamics provide a rationale for reduced copper transport ability.

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Copper relay path through the N-terminus of Wilson disease protein, ATP7B

Abstract: Using a yeast assay, we identified the roles of ATP7B's six metal-binding domains in internal copper transport and soluble chaperone capacity.

Cited by 21 publications

References 42 publications

An EPR Study on the Interaction between the Cu(I) Metal Binding Domains of ATP7B and the Atox1 Metallochaperone

An EPR Study on the Interaction between the Cu(I) Metal Binding Domains of ATP7B and the Atox1 Metallochaperone

The novel leishmanial Copper P-type ATPase plays a vital role in intracellular parasite survival

Wilson disease missense mutations in ATP7B affect metal-binding domain structural dynamics

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