Trans-2-[4-(dimethylamino)styryl]-3-ethyl-1,3-benzothiazolium perchlorate - New fluorescent dye for testing of amyloid fibrils and study of their structure

Sulatskaya, Anna I.; Sulatsky, Maksim I.; Povarova, Olga I.; Rodina, Natalia P.; Kuznetsova, Irina М.; Луговский, А. А.; Voropay, Е. S.; Lavysh, A. V.; Маскевич, А. А.; Turoverov, Konstantin К.

doi:10.1016/j.dyepig.2018.05.006

Cited by 14 publications

(16 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The last remark is important, since ThT fluorescent spectroscopy is widely used for determining the dye-fibrils binding parameters that can be used for analyzing of their structural features (see, for example, the review by Groenning [ 42 ]). In our previous works, we showed that for the solving of the problem of ThT-fibrils affinity and stoichiometry determination absorption spectroscopy of specially prepared by equilibrium microdialysis samples can be used [ 38 , 43 , 44 ].…”

Section: Resultsmentioning

confidence: 99%

“…This method was originally developed to study the interaction of low molecular weight ligands with their receptors; however, it has been undeservedly forgotten. In our recent works, we showed how this method (details of the experiment see in the Materials and Methods section) can be used for the determination of the ThT-amyloid fibrils binding parameters on the example of lysozyme and insulin fibrils [ 43 , 44 ]. It was assumed that the affinity and stoichiometry of the ThT binding to β2M amyloid fibrils could also be determined using this approach.…”

Section: Resultsmentioning

confidence: 99%

“…At the same time, the fluorescence intensity of ThT in the presence of ΔN6β2m and ΔN10β2m fibrils is comparable with the dye fluorescence in the absence of amyloid fibrils. Obtained results show that the use of equilibrium microdialysis is a key point in the samples preparation for the study of ThT interaction with ΔN6β2m and ΔN10β2m amyloid fibrils because of the large contribution of the free dye fluorescence intensity to the total fluorescence intensity (in contrast to situations in which the dye binding to fibrils is accompanied by a fluorescence intensity increase of several orders of magnitude, as in the case of β2m, lysozyme and insulin fibrils [ 43 , 44 ]).…”

Section: Resultsmentioning

confidence: 99%

“…We believe that the detected binding mode of ThT to β2M and other amyloid fibrils corresponds to the incorporation of the dye into the grooves that are formed by the amino acids side chains of amyloid protofibrils, with the dye binding along the long axis of the fibrils that is perpendicular to the β-sheets (the model proposed by Krebs [ 47 ]). However, along with this binding mode for amyloid fibrils formed from insulin and lysozyme another binding type (with ThT binding constant ~10 −6 M −1 ) was observed [ 43 , 44 ]. In order to make an assumption about the nature of the mode with high affinity in insulin and lysozyme fibrils and the reasons of its absence in β2M fibrils the morphology of these amyloid aggregates was compared.…”

Section: Resultsmentioning

confidence: 99%

See 3 more Smart Citations

Structural Features of Amyloid Fibrils Formed from the Full-Length and Truncated Forms of Beta-2-Microglobulin Probed by Fluorescent Dye Thioflavin T

Sulatskaya

Rodina

Polyakov

et al. 2018

IJMS

View full text Add to dashboard Cite

The persistence of high concentrations of beta-2-microglobulin (β2M) in the blood of patients with acute renal failure leads to the development of the dialysis-related amyloidosis. This disease manifests in the deposition of amyloid fibrils formed from the various forms of β2M in the tissues and biological fluids of patients. In this paper, the amyloid fibrils formed from the full-length β2M (β2m) and its variants that lack the 6 and 10 N-terminal amino acids of the protein polypeptide chain (ΔN6β2m and ΔN10β2m, respectively) were probed by using the fluorescent dye thioflavin T (ThT). For this aim, the tested solutions were prepared via the equilibrium microdialysis approach. Spectroscopic analysis of the obtained samples allowed us to detect one binding mode (type) of ThT interaction with all the studied variants of β2M amyloid fibrils with affinity ~104 M−1. This interaction can be explained by the dye molecules incorporation into the grooves that were formed by the amino acids side chains of amyloid protofibrils along the long axis of the fibrils. The decrease in the affinity and stoichiometry of the dye interaction with β2M fibrils, as well as in the fluorescence quantum yield and lifetime of the bound dye upon the shortening of the protein amino acid sequence were shown. The observed differences in the ThT-β2M fibrils binding parameters and characteristics of the bound dye allowed to prove not only the difference of the ΔN10β2m fibrils from other β2M fibrils (that can be detected visually, for example, by transmission electron microscopy (TEM), but also the differences between β2m and ΔN6β2m fibrils (that can not be unequivocally confirmed by other approaches). These results prove an essential role of N-terminal amino acids of the protein in the formation of the β2M amyloid fibrils. Information about amyloidogenic protein sequences can be claimed in the development of ways to inhibit β2M fibrillogenesis for the treatment of dialysis-related amyloidosis.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Structural Features of Amyloid Fibrils Formed from the Full-Length and Truncated Forms of Beta-2-Microglobulin Probed by Fluorescent Dye Thioflavin T

Sulatskaya

Rodina

Polyakov

et al. 2018

IJMS

View full text Add to dashboard Cite

show abstract

“…In addition, the interaction of DMASEBT with amyloid fibrils formed from insulin was investigated and it was shown that the photophysical characteristics of the analogue significantly change (similarly in the case of ThT) when the dye binds to these fibrils compared to the characteristics of the free DMASEBT in aqueous solution. 15 In the present work, the study of DMASEBT interaction with amyloid fibrils was continued. Using the specially elaborated approach based on the spectroscopic study of solutions prepared by the equilibrium microdialysis, 18−22 the parameters of the dye binding to amyloid fibrils formed from lysozyme and photophysical properties of the bound to the fibrils dye were determined.…”

mentioning

confidence: 91%

Structural Analogue of Thioflavin T, DMASEBT, as a Tool for Amyloid Fibrils Study

et al. 2019

View full text Add to dashboard Cite

Fluorescent dye trans-2-[4-(dimethylamino)styryl]-3-ethyl-1,3-benzothiazolium perchlorate (DMASEBT) is a relatively recently synthesized probe for detection of amyloid fibrils accumulating in the organs and tissues of patients with a wide range of serious incurable diseases. DMASEBT was developed as an alternative of its widely used analogue thioflavin T (ThT), which is the "gold standard" for the amyloid fibrils study. Our results show the similarity of both dyes binding to amyloid fibrils and allow one to propose a mechanism of such probes interaction with some types of the fibrils. At the same time, DMASEBT has a significant advantage, namely, improved photophysical properties compared with ThT, which allows for the detection of DMASEBT-stained amyloid fibrils in the spectral region of the "transparency window of biological tissues". The ability of the dye to penetrate into the cells was shown to open the prospect of this dye's use for amyloid fibrils bioimaging and biosensing in vivo. Furthermore, it was proven that DMASEBT can be used not only as a test for amyloid fibrils formation but also for the comparative study of the fibrils structure (both their fibers and bunches), which in turn may underlie the variability of amyloidosis and affect the cytotoxicity of these protein aggregates.

show abstract

Luminescence Spectral Properties of New Benzothiazole Polymethine Dye

et al. 2023

View full text Add to dashboard Cite

The absorption and fl uorescence spectra of a new styryl derivative of thiofl avin T 2-{(1E,3E)-4- in solvents with diff erent polarity and viscosity and also incorporated in the structure of amyloid fi brils and bovine serum albumin were investigated. A characteristic feature of the dye is an extremely low quantum yield of fl uorescence in low-viscosity solvents (10 -4 in water) which, however, increases signifi cantly in viscous solutions and when it is incorporated in the structure of proteins or amyloid fi brils. In the latter case the quantum yield increases by 8•10 3 times. On the basis of the experimental studies and quantum chemical calculations it was shown that Th-C23 exhibits the properties of a molecular rotor. The increase of the fl uorescence quantum yield in viscous solutions and in the biopolymers results from limitation of the torsional rotation of the molecular fragments, leading to fl uorescence quenching. The longwavelength location of the absorption spectrum and the fl uorescence spectrum of the new dye in the red region of the spectrum (520 and 600 nm) makes it possible to use it as a fl uorescent marker that is sensitive to the viscosity (hardness) of the microenvironment not only in vitro but also in vivo.

show abstract

Trans-2-[4-(dimethylamino)styryl]-3-ethyl-1,3-benzothiazolium perchlorate - New fluorescent dye for testing of amyloid fibrils and study of their structure

Cited by 14 publications

References 35 publications

Structural Features of Amyloid Fibrils Formed from the Full-Length and Truncated Forms of Beta-2-Microglobulin Probed by Fluorescent Dye Thioflavin T

Structural Features of Amyloid Fibrils Formed from the Full-Length and Truncated Forms of Beta-2-Microglobulin Probed by Fluorescent Dye Thioflavin T

Structural Analogue of Thioflavin T, DMASEBT, as a Tool for Amyloid Fibrils Study

Luminescence Spectral Properties of New Benzothiazole Polymethine Dye

Contact Info

Product

Resources

About