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Updated: Jul 3, 2026

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
Published on: December 16, 2013
Conserved residues modulate copper release in human copper chaperone Atox1
Faiza Hussain1, John S Olson, Pernilla Wittung-Stafshede
1Department of Biochemistry and Cell Biology, Keck Center for Structural Computational Biology, Houston, TX 77251, USA.
The human copper chaperone Atox1 facilitates copper delivery to Wilson and Menkes disease proteins. Mutants showed faster copper release, suggesting chaperones enhance copper accessibility for disease protein targets.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Copper (Cu) chaperones like Atox1 are crucial for intracellular copper transport.
- Defects in copper transport are linked to neurodegenerative diseases such as Wilson disease and Menkes disease.
- The precise mechanism of copper delivery from Atox1 to its target proteins in the cytoplasm remains poorly understood.
Purpose of the Study:
- To investigate the mechanism of copper(I) release from the human copper chaperone Atox1.
- To characterize the role of specific residues (Met10 and Lys60) in copper binding and release.
- To compare the kinetics of copper transfer from Atox1 to a chelator with copper uptake from solution.
Main Methods:
- Characterization of copper(I) release from wild-type Atox1 and point mutants (Met10Ala, Lys60Ala).
- Utilized bicinchoninic acid (BCA) as a copper(I) chelator and metal acceptor.
- Measured the dynamics of copper(I) displacement using a three-step kinetic model.
Main Results:
- Copper(I) displacement from holo-Atox1 by BCA follows a three-step process involving complex formation and dissociation.
- Both Met10Ala and Lys60Ala mutants released copper(I) more readily than wild-type Atox1.
- Copper(I) transfer from holo-Atox1 to BCA was significantly faster than direct copper(I) uptake by BCA from solution.
Conclusions:
- Copper chaperones like Atox1 play a vital role in rapidly making copper(I) available to cellular substrates.
- The activated protein-metal-chelator complex may kinetically mimic the in vivo ternary chaperone-metal-target complex.
- Atox1 mutations affecting copper release kinetics provide insights into copper transport mechanisms relevant to Wilson and Menkes diseases.
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