Ultrafast torsional dynamics of Thioflavin-T in an anionic cyclodextrin cavity

Singh, Prabhat; Murudkar, Sushant; Mora, Aruna K.; Nath, Sukhendu

doi:10.1016/j.jphotochem.2014.10.007

Cited by 46 publications

(51 citation statements)

References 57 publications

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“…When ThT is bound to an amyloid fibril or found in other confining solvents (e.g. SDS micelles [88], high viscosity solvents [89][90][91], or cyclodextrin [92]), the inter-ring rotation is slowed down, favoring highly fluorescent relaxation directly from the LE to the ground state and increasing fluorescent yields by factors ranging from 160 in amyloid [86] to 13 in SDS [88], in comparison to the free dye. Decreasing solvent temperatures will increase the inherent ThT fluorescence due to increased solvent viscosity [89,91].…”

Section: Viscosity and The Molecular Rotormentioning

confidence: 99%

“…This does not necessarily rule out the use of ThT to report on the following fibrillation phase, as long as the following kinetic trace is robust and well above the elevated baseline. Examples of exogenous compounds that can elevate ThT emission include SDS [88], b-cyclodextrin [92], and glycosaminoglycans (Malmos et al unpublished results). Fibrilmediated interactions between compounds and ThT as well as false negatives (samples containing amyloid fibrils but showing no ThT emission) are more difficult to detect and must be complemented by other assays (check for more explanation in following paragraphs).…”

Section: In Situ Vs Fixed Time Point Assaysmentioning

confidence: 99%

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ThT 101: a primer on the use of thioflavin T to investigate amyloid formation

Malmos

Blancas‐Mejia

Weber

et al. 2017

Amyloid

322

243

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Thioflavin T (ThT) has been widely used to investigate amyloid formation since 1989. While concerns have recently been raised about its use as a probe specific for amyloid, ThT still continues to be a very valuable tool for studying kinetic aspects of fibrillation and associated inhibition mechanisms. This review aims to provide a conceptual instruction manual, covering appropriate considerations and pitfalls related to the use of ThT. We start by giving a brief introduction to amyloid formation with focus on the morphology of different aggregate species, followed by a discussion of the quality of protein needed to obtain reliable fibrillation data. After an overview of the photochemical basis for ThT's amyloid binding properties and artifacts that may arise from this, we describe how to plan and analyze ThT assays. We conclude with recommendations for complementary techniques to address shortcomings in the ThT assay.

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Section: Viscosity and The Molecular Rotormentioning

confidence: 99%

Section: In Situ Vs Fixed Time Point Assaysmentioning

confidence: 99%

ThT 101: a primer on the use of thioflavin T to investigate amyloid formation

Malmos

Blancas‐Mejia

Weber

et al. 2017

Amyloid

322

243

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“…Solution fluorometric titration of ThT with PβCD in 50 m m HEPES buffer (Figure S4, SI) yielded a dissociation constant of 3.25±0.14 μ m for the complex, assuming a 1:1 binding model. This is 30‐fold lower (i.e., stronger binding) than for complexes between ThT and sulfobutylether βCD in deionized water …”

Section: Methodsmentioning

confidence: 86%

“…Negative ion mode nanoESI‐MS of an aqueous solution of 25 μ m ThT and 25 μ m PβCD shows that 1:1 binding was favored in complexes with net 2− charge state. In contrast, complexes between ThT and non‐derivatized βCD ( K d =13 m m ) were not detected using nanoESI mass spectrometry in either positive or negative ion modes (Figure S3, SI). Solution fluorometric titration of ThT with PβCD in 50 m m HEPES buffer (Figure S4, SI) yielded a dissociation constant of 3.25±0.14 μ m for the complex, assuming a 1:1 binding model.…”

Section: Methodsmentioning

confidence: 99%

“…Our choice to examine ThT·PbCD complexes was inspired by the knowledge that CDs with extended cavities facilitate bindingo fs mall molecules, [23] and that a % 17-fold fluorescence enhancement occurs when ThT is boundt oC aptisol, a polyanionic bCD modifiedw ith sulfonates tethered by butylether linkages. [24] Moreover,T hT·PbCD complexes seemed likely to survive desolvation because of favorable electrostatic interactions between cationic ThT and anionic carboxylate tails of the extended bCD cavity. Figure 1s hows that ThT binds well to PbCD and that the complex is more fluorescent than ThT + alone in both the gas and solution phases.…”

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confidence: 99%

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Intrinsic Turn‐On Response of Thioflavin T in Complexes

Kung

Vurgun

Chen

et al. 2020

Chemistry A European J

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The fluorescence enhancement (“turn‐on”) response of the amyloid‐sensing dye thioflavin T (ThT) is examined in vacuo, where solvent interactions are absent. Upon the complexation of ThT with a derivatized β‐cyclodextrin, heptakis‐[6‐deoxy‐6‐(3‐sulfanylpropanoic acid)]‐β‐cyclodextrin, turn‐on responses in both the gas phase and solution phase were observed. In contrast, turn‐on response was not detected when ThT was bound to gaseous cucurbit[7]uril or human telomeric DNA 22AG, whereas clear turn‐on response occurs in solution. The observed difference in turn‐on response in the gas phase emphasizes the key interplay between chromophore, host and solvent and demonstrates the utility of gas‐phase spectroscopy to tease out the balance among intermolecular forces driving the behavior of important chromophores in solution.

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Proton Transfer Reaction Dynamics of Pyranine in DMSO/Water Mixtures

Awasthi¹,

Singh

2017

ChemPhysChem

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Photoinduced intermolecular excited-state proton transfer (ESPT) reactions are ubiquitous in chemistry and biology. ESPT reactions are extremely sensitive to the nature of water molecules in its microenvironment and thus serve as a sensitive reporter for the water structure and dynamics in a system. Herein, the photoinduced intermolecular ESPT reaction of 8-hydroxypyrene-1,3,5-trisulfonic acid (HPTS, also known as pyranine) has been investigated in various DMSO/water mixtures by using steady-state and time-resolved emission spectroscopy. The DMSO/water binary mixture yields an interesting and anomalous behavior for the proton transfer reaction dynamics of HPTS at a mole fraction of DMSO (X ) of 0.41-0.51, which has also been previously investigated and projected as an anomalous region by molecular dynamics simulation and other experimental techniques. The extreme slowdown of the proton transfer reaction observed at X =0.41-0.51 has been attributed to the slow solvation dynamics, as well as the non-availability of free water molecules at this composition, which are required to solvate the newly generated proton. These observations have been also corroborated by time-resolved area-normalized emission spectra. The dimensionality of the proton diffusion process has been investigated by analyzing the geminate recombination process, and is found to be significantly different in DMSO/water mixtures (X =0.41-0.51) compared with three-dimensional proton diffusion in neat water.

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Ultrafast torsional dynamics of Thioflavin-T in an anionic cyclodextrin cavity

Cited by 46 publications

References 57 publications

ThT 101: a primer on the use of thioflavin T to investigate amyloid formation

ThT 101: a primer on the use of thioflavin T to investigate amyloid formation

Intrinsic Turn‐On Response of Thioflavin T in Complexes

Proton Transfer Reaction Dynamics of Pyranine in DMSO/Water Mixtures

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