Probing transient copper Chaperone-Wilson disease protein interactions at the single-molecule level with nanovesicle trapping

Jaime J. Benítez, Aaron M. Keller, Patrick Ochieng, Liliya A. Yatsunyk, David L. Huffman*, Amy C. Rosenzweig, Peng Chen

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2446-2447
Number of pages2
JournalJournal of the American Chemical Society
Volume130
Issue number8
DOIs
StatePublished - Feb 27 2008

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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