Wilson's Disease Research Today is a free monthly online journal that collates and summarizes the latest research about Wilson's Disease, including details on treatment, causes, symptoms, medication.

Probing Transient Copper Chaperone-Wilson Disease Protein Interactions at the Single-Molecule Level with Nanovesicle Trapping.

Benítez JJ, Keller AM, Ochieng P, Yatsunyk LA, Huffman DL, Rosenzweig AC, Chen P

Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, Department of Chemistry, Western Michigan University, Kalamazoo, Michigan 49008, and Departments of Biochemistry, Molecular Biology, and Cell Biology and of Chemistry, Northwestern University, Evanston, Illinois 60208.

Transient metallochaperone-target protein interactions are essential for intracellular metal trafficking but challenging to study at both the ensemble and the single-molecule level. Here we report using nanovesicle trapping to enable single-molecule fluorescence resonance energy transfer (smFRET) studies of transient interactions between the copper chaperone Hah1 and the fourth metal-binding domain of its target protein, the Wilson disease protein (WDP). We were able to monitor their interactions in real time one event at a time, capture distinct protein interaction intermediates, resolve intermediate interconversion dynamics, and quantify both the interaction kinetics and thermodynamics in the absence of copper. The study exemplifies the ability of nanovesicle trapping in combination with smFRET for studying weak protein interactions and provides insight into how Hah1 and WDP may collaborate to mediate copper transfer inside cells.

Published 5 February 2008 in J Am Chem Soc.
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