scholarly journals The Fluorescent Dye 1,6-Diphenyl-1,3,5-hexatriene Binds to Amyloid Fibrils Formed by Human Amylin and Provides a New Probe of Amylin Amyloid Kinetics

Biochemistry ◽  
2021 ◽  
Author(s):  
Ming-Hao Li ◽  
Lakshan Manathunga ◽  
Erwin London ◽  
Daniel P. Raleigh
Keyword(s):  
Amyloid Fibrils ◽  
Fluorescent Dye ◽  
Human Amylin

scholarly journals The Fluorescent Dye 1,6-Diphenyl-1,3,5-Hexatriene Binds to Amyloid Fibrils Formed by Human Amylin and Provides a New Probe of Amylin Amyloid Kinetics

2021 ◽  
Author(s):  
Ming-Hao Li ◽  
Lakshan Manathunga ◽  
Erwin London ◽  
Daniel Raleigh
Keyword(s):  
Time Course ◽  
Amyloid Fibrils ◽  
Fold Increase ◽  
Fluorescent Dye ◽  
Thioflavin T ◽  
Lag Phase ◽  
Amyloid Formation ◽  
Islet Amyloid ◽  
Human Amylin ◽  
Mutant Forms

The fluorescent dye 1,6-diphenyl-1,3,5-hexatriene (DPH) is widely used as a probe of membrane order. We show that DPH also interacts with amyloid fibrils formed by human amylin (also known as islet amyloid polypeptide) in solution and this results in a 100-fold increase in DPH fluorescence for a sample of microM human amylin and 0.25 microM DPH. No increase in DPH fluorescence is observed with the non-amyloidogenic rat amylin or with freshly dissolved, non-fibrillar human amylin. The time course of amyloid formation by amylin was followed by monitoring the fluorescence of added DPH as a function of time and was similar to that monitored by the standard fluorescent probe thioflavin-T. The inclusion of DPH in the buffer did not perturb the time course of amyloid formation under the conditions examined and the time course was independent of the range of DPH concentrations tested (0.25 to 5 microM). Maximum final fluorescence intensity is observed at substoichiometric ratios of DPH to amylin. No significant increase in fluorescence was observed during the lag phase of amyloid formation, and the implications for the structure of amylin pre-fibril oligomers are discussed. Human amylin contains three aromatic residues. A triple aromatic to leucine mutant forms amyloid and DPH binds to the resulting fibrils, indicating that interactions with aromatic side chains are not required for DPH amylin amyloid interactions. DPH may be especially useful for studies on mutant amylins and other polypeptides in which changes in charged residues might complicate interpretation of thioflavin-T fluorescence.

Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavine T

1989 ◽  
Vol 177 (2) ◽  
pp. 244-249 ◽  
Author(s):  
Hironobu Naiki ◽  
Keiichi Higuchi ◽  
Masanori Hosokawa ◽  
Toshio Takeda
Keyword(s):  
Amyloid Fibrils ◽  
Fluorescent Dye ◽  
Fluorometric Determination ◽  
Thioflavine T

The Parallel Superpleated Beta-structure as a Model for Amyloid Fibrils of Human Amylin

2005 ◽  
Vol 348 (2) ◽  
pp. 247-252 ◽  
Author(s):  
Andrey V. Kajava ◽  
Ueli Aebi ◽  
Alasdair C. Steven
Keyword(s):  
Amyloid Fibrils ◽  
Beta Structure ◽  
Human Amylin

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

2018 ◽  
Vol 19 (9) ◽  
pp. 2762 ◽  
Author(s):  
Anna Sulatskaya ◽  
Natalia Rodina ◽  
Dmitry Polyakov ◽  
Maksim Sulatsky ◽  
Tatyana Artamonova ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amyloid Fibrils ◽  
Biological Fluids ◽  
Fluorescent Dye ◽  
Full Length ◽  
Thioflavin T ◽  
Protein Amino Acid ◽  
Terminal Amino ◽  
Beta 2 ◽  
Beta 2 Microglobulin

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.

Spectroscopic characterization of diverse amyloid fibrils in vitro by the fluorescent dye Nile red

2011 ◽  
Vol 7 (4) ◽  
pp. 1232 ◽  
Author(s):  
Rajesh Mishra ◽  
Daniel Sjölander ◽  
Per Hammarström
Keyword(s):  
Amyloid Fibrils ◽  
Nile Red ◽  
Spectroscopic Characterization ◽  
Fluorescent Dye

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

2018 ◽  
Vol 157 ◽  
pp. 385-395 ◽  
Author(s):  
Anna I. Sulatskaya ◽  
Maksim I. Sulatsky ◽  
Olga I. Povarova ◽  
Natalia P. Rodina ◽  
Irina M. Kuznetsova ◽  
...  
Keyword(s):  
Amyloid Fibrils ◽  
Fluorescent Dye

scholarly journals The toxic nature of murine amylin and the immune responsivity of pancreatic islet to conformational antibody in mice

2017 ◽  
Author(s):  
Luiza C. S. Erthal ◽  
Luana Jotha-Mattos ◽  
Flávio Alves Lara ◽  
Sabrina Alves dos Reis ◽  
Bernardo Miguel de Oliveira Pascarelli ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Pancreatic Islet ◽  
Amyloid Fibrils ◽  
Peptide Hormone ◽  
List Type ◽  
Pancreatic Peptide ◽  
Human Amylin ◽  
First Time

ABSTRACTThe human amylin is a pancreatic peptide hormone cosecreted with amylin and found in hyperhormonemic state along with insulin in subclinical diabetes. Amylin has been associated with the pathology of type 2 diabetes, particularly due to its ability to assembly into toxic oligomers and amyloid speciments. On the other hand, some variants such as murine amylin has been described as non amyloidogenic, either in vitro or in vivo. Recent data have demonstrated the amyloid propensity of murine amylin and the therapeutic analogue pramlintide, suggesting a universality for amylin amyloidosis. Here we report the amyloidogenesis of murine amylin, which showed lower responsivity to the fluorescent probe thioflavin T compared to human amylin, but presented highly organized fibrilar amyloid material. The aggregation of murine amylin also resulted in the formation of cytotoxic specimens, as evaluated in vitro in INS-1 cells. The aggregation product from murine amylin was responsive to a specific antibody raised against amyloid oligomers, the A11 oligomer antibody. Pancreatic islets of swiss mice have also shown responsivity for the anti-oligomer, indicating the natural abundance of such specimen in rodents. These data provide for the first time evidences for the toxic nature of oligomeric assemblies of murine amylin and its existence in non-transgenic mice.Highlights- Murine amylin forms oligomer species and amyloid fibrils in vitro- The murine amylin aggregation product display cellular toxicity- A11 anti-oligomer antibody recognizes murine amylin in vitro- Non-transgenic mice display immunoresposivity to anti-oligomer in pancreatic islet

Liquid crystalline ordering of paired helical filaments in neurofibrillary tangles of Alzheimer's disease

1986 ◽  
Vol 44 ◽  
pp. 348-349
Author(s):  
D.F. Clapin ◽  
V.J.A. Montpetit
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Neurofibrillary Tangles ◽  
Liquid Crystalline ◽  
Amyloid Fibrils ◽  
Geometric Analysis ◽  
Paired Helical Filaments ◽  
Microscopic Level ◽  
Light Microscopic Level ◽  
Regular Spacing

Alzheimer's disease is characterized by the accumulation of abnormal filamentous proteins. The most important of these are amyloid fibrils and paired helical filaments (PHF). PHF are located intraneuronally forming bundles called neurofibrillary tangles. The designation of these structures as "tangles" is appropriate at the light microscopic level. However, localized domains within individual tangles appear to demonstrate a regular spacing which may indicate a liquid crystalline phase. The purpose of this paper is to present a statistical geometric analysis of PHF packing.

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