Blue fluorescent dye-protein complexes based on fluorogenic cyanine dyes and single chain antibody fragments

Zanotti, Kimberly J.; Silva, Gloria L.; Creeger, Yehuda; Robertson, Kelly L.; Waggoner, Alan S.; Berget, Peter B.; Armitage, Bruce A.

doi:10.1039/c0ob00444h

Cited by 57 publications

(66 citation statements)

References 27 publications

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“…3,4,6,121 The radiationless decay of monomethine dyes in condensed phase has been studied as an example of a barrierless viscosity-controlled process. 21,28,[39][40][41] Predictions of internal charge-transfer coupled to the twisting motion have been known for some time.…”

Section: A General Discussionmentioning

confidence: 99%

A two-state model of twisted intramolecular charge-transfer in monomethine dyes

Olsen

McKenzie

2012

The Journal of Chemical Physics

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Articles you may be interested inModeling opto-electronic properties of a dye molecule in proximity of a semiconductor nanoparticle J. Chem. Phys. 139, 024105 (2013) A two-state model Hamiltonian is proposed, which can describe the coupling of twisting displacements to charge-transfer behavior in the ground and excited states of a general monomethine dye molecule. This coupling may be relevant to the molecular mechanism of environment-dependent fluorescence yield enhancement. The model is parameterized against quantum chemical calculations on different protonation states of the green fluorescent protein chromophore, which are chosen to sample different regimes of detuning from the cyanine (resonant) limit. The model provides a simple yet realistic description of the charge transfer character along two possible excited state twisting channels associated with the methine bridge. It describes qualitatively different behavior in three regions that can be classified by their relationship to the resonant (cyanine) limit. The regimes differ by the presence or absence of twist-dependent polarization reversal and the occurrence of conical intersections. We find that selective biasing of one twisting channel over another by an applied diabatic biasing potential can only be achieved in a finite range of parameters near the cyanine limit.

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Section: A General Discussionmentioning

confidence: 99%

A two-state model of twisted intramolecular charge-transfer in monomethine dyes

Olsen

McKenzie

2012

The Journal of Chemical Physics

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“…Furthermore, in order to increasingly accelerate the off-rates of signal detection using multiselective FAPs, the future focus will be placed on isolating weakaffinity fluorogens. Here, two distinct engineering strategies may rapidly facilitate this: the site-directed mutagenesis of the protein scaffold or chemical alterations of fluorogen sub-groups (Rastede et al, 2015;Shank et al, 2009;Yates et al, 2013;Zanotti et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…Determining multi-fluorogen activation from scFv HL1.0.1-TO1 and HL1-TO1 at the surface of cells Based on our fluorescence validation screen, we selected three different spectra cell-impermeant fluorogens for further validation on live cells (Fig. S4): a Oxazole Thiazole Blue derivative (OTB-SO 3 ) (Zanotti et al, 2011), a Thiazole Orange derivative (TO1-2p), and Dimethyl-Indole Red (DIR) (Constantin et al, 2008). In addition, we included in our assessment scFv HL1.0.1-TO1 and its parent scFv HL1-TO1 (a non-affinity matured version) (SzentGyorgyi et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Breaking the color barrier – a multi-selective antibody reporter offers innovative strategies of fluorescence detection

Gallo

Jarvik

2017

Journal of Cell Science

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A novel bi-partite fluorescence platform exploits the high affinity and selectivity of antibody scaffolds to capture and activate small-molecule fluorogens. In this report, we investigated the property of multiselectivity activation by a single antibody against diverse cyanine family fluorogens. Our fluorescence screen identified three cellimpermeant fluorogens, each with unique emission spectra (blue, green and red) and nanomolar affinities. Most importantly, as a protein fusion tag to G-protein-coupled receptors, the antibody biosensor retained full activity -displaying bright fluorogen signals with minimal background on live cells. Because fluorogen-activating antibodies interact with their target ligands via non-covalent interactions, we were able to perform advanced multi-color detection strategies on live cells, previously difficult or impossible with conventional reporters. We found that by fine-tuning the concentrations of the different color fluorogen molecules in solution, a user may interchange the fluorescence signal (onset versus offset), execute real-time signal exchange via fluorogen competition, measure multi-channel fluorescence via colabeling, and assess real-time cell surface receptor traffic via pulsechase experiments. Thus, here we inform of an innovative reporter technology based on tri-color signal that allows user-defined fluorescence tuning in live-cell applications.

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“…This property has been exploited to create membrane-permeant and -impermeant fluorogens, amplify fluoromodule brightness, and build activatable pH-reporting ratiometric probes that can all be activated by the same FAP (20,(22)(23)(24). All previously described fluoromodules, with one exception (K7:TO-PRO-5), absorb and fluoresce maximally in the visible light spectrum, which limits their effectiveness for animal imaging experiments (20,(25)(26)(27)(28).…”

Section: Introductionmentioning

confidence: 99%