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Copper-mediated homo-dimerisation for the HAH1 metallochaperone
Valérie Tanchou1, Fabienne Gas, Agathe Urvoas
1CEA VALRHO, Département d'Ingénierie et d'Etudes des Protéines, Service de Biochimie Post-génomique et Toxicologie Nucléaire, BP1717, 30207 Bagnols sur Cèze cedex, France. valerie.tanchou@cea.fr
Biochemical and Biophysical Research Communications
|November 9, 2004
Summary
The HAH1 metallochaperone forms dimers with copper, interacting with Menkes ATPase. This dimerization, confirmed in cells, is crucial for intracellular copper control.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Copper is essential for human cells, and its homeostasis is tightly regulated.
- The HAH1 metallochaperone plays a critical role in managing intracellular copper levels.
- Dysregulation of copper homeostasis is linked to various diseases.
Purpose of the Study:
- To investigate the role of copper in HAH1 metallochaperone function.
- To elucidate the interaction between HAH1 and the Menkes ATPase.
- To confirm HAH1 homo-dimerization in a cellular context.
Main Methods:
- Solid-phase based assay
- Biacore analysis for kinetic parameter determination
- Fluorescence resonance energy transfer (FRET) in living cells
Main Results:
- HAH1 forms stable homo-dimers in the presence of copper, with an apparent affinity constant of 6µM.
- Copper-loaded HAH1 interacts independently with the six metal-binding domains of Menkes ATPase.
- HAH1 homo-dimerization was validated in living cells using FRET.
Conclusions:
- Copper binding induces HAH1 homo-dimerization, a key step in its function.
- The HAH1-Menkes ATPase interaction is modular, involving distinct binding domains.
- These findings provide insights into the mechanisms of intracellular copper regulation by HAH1.