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

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.

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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 ◽

10.1021/acs.biochem.1c00328 ◽

2021 ◽

Author(s):

Ming-Hao Li ◽

Lakshan Manathunga ◽

Erwin London ◽

Daniel P. Raleigh

Keyword(s):

Amyloid Fibrils ◽

Fluorescent Dye ◽

Human Amylin

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The triphenylmethane dye brilliant blue G is only moderately effective at inhibiting amyloid formation by human amylin or at disaggregating amylin amyloid fibrils, but interferes with amyloid assays; Implications for inhibitor design

PLoS ONE ◽

10.1371/journal.pone.0219130 ◽

2019 ◽

Vol 14 (8) ◽

pp. e0219130

Author(s):

Rehana Akter ◽

Alexander Zhyvoloup ◽

Bingqian Zheng ◽

Surita R. Bhatia ◽

Daniel P. Raleigh

Keyword(s):

Amyloid Fibrils ◽

Brilliant Blue ◽

Amyloid Formation ◽

Inhibitor Design ◽

Brilliant Blue G ◽

Triphenylmethane Dye ◽

Human Amylin

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

10.1101/175539 ◽

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

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Crystalline order to a resolution of 4.7 A in the sheath of Methanospirilium hungatii: A cross-beta structure

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111185 ◽

1984 ◽

Vol 42 ◽

pp. 214-215

Author(s):

Murray Stewart ◽

T.J. Beveridge ◽

D. Sprott

Keyword(s):

Cell Wall ◽

Single Layer ◽

Uranyl Acetate ◽

Optical Diffraction ◽

Tube Axis ◽

Basic Subunit ◽

Beta Structure ◽

Diffraction Patterns ◽

Protein Treatment ◽

General Appearance

The archaebacterium Methanospirillum hungatii has a sheath as part of its cell wall which is composed mainly of protein. Treatment with dithiothreitol or NaOH released the intact sheaths and electron micrographs of this material negatively stained with uranyl acetate showed flattened hollow tubes, about 0.5 μm diameter and several microns long, in which the patterns from the top and bottom were superimposed. Single layers, derived from broken tubes, were also seen and were more simply analysed. Figure 1 shows the general appearance of a single layer. There was a faint axial periodicity at 28.5 A, which was stronger at irregular multiples of 28.5 A (3 and 4 times were most common), and fine striations were also seen at about 3° to the tube axis. Low angle electron diffraction patterns (not shown) and optical diffraction patterns (Fig. 2) from these layers showed a complex meridian (as a result of the irregular nature of the repeat along the tube axis) which showed a clear maximum at 28.5 A, consistent with the basic subunit spacing.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100143365 ◽

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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