2008
DOI: 10.1021/ja7107867
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Probing Transient Copper Chaperone−Wilson Disease Protein Interactions at the Single-Molecule Level with Nanovesicle Trapping

Abstract: Metals are essential nutrients that can also be toxic. Safe trafficking of metal ions is necessary inside cells, and specific metal transport pathways exist to deliver them to their destinations. 1,2 In human cells, the copper chaperone Hah1 and the Wilson disease protein (WDP) constitute a copper transport pathway-Hah1 is a single-domain cytoplasmic protein; WDP is a multidomain protein anchored on organelle membranes and has a cytosolic N-terminal region consisting of six homologous metal-binding domains (MB… Show more

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Cited by 50 publications
(77 citation statements)
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“…Following a similar reasoning, researchers have been able to capture fluorescently labeled binding partners in small (100-200 nm), surface-tethered lipid nanovesicles and study their interactions through single-molecule FRET measurements (Figure 2b) [51][52][53]54 ]. The volume of a 100-nm vesicle is two orders of magnitude smaller than that of a diffraction-limited volume and thus allows proteins to be visualized at the single-molecule level at much higher effective concentrations than hitherto possible.…”
Section: The Concentration Problemmentioning
confidence: 98%
“…Following a similar reasoning, researchers have been able to capture fluorescently labeled binding partners in small (100-200 nm), surface-tethered lipid nanovesicles and study their interactions through single-molecule FRET measurements (Figure 2b) [51][52][53]54 ]. The volume of a 100-nm vesicle is two orders of magnitude smaller than that of a diffraction-limited volume and thus allows proteins to be visualized at the single-molecule level at much higher effective concentrations than hitherto possible.…”
Section: The Concentration Problemmentioning
confidence: 98%
“…A very attractive alternative is to restrict individual molecules within small, optically transparent containers. Examples include lipid vesicles (20)(21)(22)(23)(24), selfassembled polymer nanocapsules (25,26), protein shells of viral capsids (27), and femtoliter chambers chemically etched into the surface of glass optical fibers (28,29).…”
mentioning
confidence: 99%
“…9a) (58). Benitez et al were able to observe two transiently bound states with different FRET ratios (E 1 ~ 0.5, E 2 ~ 0.9) that can interconvert and quantify all rate constants for the binding scheme (Fig.…”
Section: Applicationsmentioning
confidence: 99%