Preparation and Characterization of Purified Amyloid Fibrils

Zurdo, Jesús; Guijarro, J. Iñaki; Dobson, Christopher M.

doi:10.1021/ja016229b

Cited by 123 publications

(117 citation statements)

References 10 publications

Supporting

Mentioning

110

Contrasting

Unclassified

Order By: Relevance

“…The amide I band (1575-1725 cm Ϫ1 ) of each spectrum was subjected to a fitting procedure using seven Gaussian distributions centered at the frequencies of well characterized secondary structures and lateral chains (27,28). Each Gaussian was characterized by its frequency and an interval of given width.…”

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Monsellier

Redeker

Ruiz-Arlandis

et al. 2015

Journal of Biological Chemistry

103

View full text Add to dashboard Cite

Background: Hsc70 has an alleviating effect on the toxicity of polyglutamine (polyQ)-containing proteins in vivo. Results: Hsc70 binds specifically the N-terminal flank of huntingtin exon 1. Conclusion: Hsc70 interaction with huntingtin exon 1 N-terminal flank affects the conformation of the resulting assemblies. Significance: We identify the surface interfaces between Hsc70 and huntingtin exon 1, which allows the design of future therapeutic tools.

show abstract

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Monsellier

Redeker

Ruiz-Arlandis

et al. 2015

Journal of Biological Chemistry

103

View full text Add to dashboard Cite

show abstract

“…1B). Prior to analysis by x-ray fiber diffraction, the ␣-crystallin fibrils were purified according to the method of Zurdo et al (22). No detectable quantities of non-fibrillar aggregates were observed in the TEM images of ␤-and ␥-crystallin fibril samples; thus, no further purification was required for these specimens.…”

Section: Fibril Formation By ␣-Crystallin At Neutral Ph-␣-crystal-mentioning

confidence: 99%

Amyloid Fibril Formation by Lens Crystallin Proteins and Its Implications for Cataract Formation

Meehan

Berry

Luisi

et al. 2004

Journal of Biological Chemistry

Self Cite

175

186

View full text Add to dashboard Cite

The ␣-, ␤-, and ␥-crystallins are the major structural proteins within the eye lens and are responsible for its exceptional stability and transparency. Under mildly denaturing conditions, all three types of bovine crystallin assemble into fibrillar structures in vitro. Characterization by transmission electron microscopy, dye binding assays, and x-ray fiber diffraction shows that these species have all of the characteristics of fibrils associated with the family of amyloid diseases. Moreover, the full-length proteins are incorporated into the fibrils, (i.e. no protein cleavage is required for these species to form), although for the ␥-crystallins some fragmentation occurs under the conditions employed in this study. Our findings indicate that the inherent stability of the ␤-sheet supramolecular structure adopted by the crystallins in the eye lens and the chaperone ability of ␣-crystallin must be crucial for preventing fibril formation in vivo. The crystallins are very stable proteins but undergo extensive post-translational modification with age that leads to their destabilization. The ability of the crystallins to convert into fibrils under destabilizing conditions suggests that this process could contribute to the development of cataract with aging.

show abstract

“…1A). The overall molecular structure of amyloid fibrils is stabilized by a dense network of H-bonds 4 , and features exceptional mechanical properties 5,6 such as high stiffness [5][6][7][8] , selfassembling capacity [9][10][11][12] , and remarkable stability even in adverse chemical environments 13 .…”

Section: Introductionmentioning

confidence: 99%

Self-folding and aggregation of amyloid nanofibrils

2011

View full text Add to dashboard Cite

Amyloids are highly organized protein filaments, rich in beta-sheet secondary structures that self-assemble to form dense plaques in brain tissues affected by severe neurodegenerative disorders (e.g. Alzheimer's Disease). Identified as natural functional materials in bacteria, in addition to their remarkable mechanical properties, amyloids have also been proposed as a platform for novel biomaterials in nanotechnology applications including nanowires, liquid crystals, scaffolds and thin films. Despite recent progress in the understanding amyloid structure and behavior, the latent selfassembly mechanism and the underlying adhesion forces that drive the aggregation process remain poorly understood. On the basis of previous full atomistic simulations, here we report a simple coarse-grain model to analyze the competition between adhesive forces and elastic deformation of amyloid fibrils. We use simple model system to investigate self-assembly mechanisms of fibrils, focused on the formation of self-folded nanorackets and nanorings, and thereby address a critical issue in linking the biochemical (Angstrom) to micrometer scales relevant for larger-scale states of functional amyloid materials. We investigate the effect of varying the interfibril adhesion energy on the structure and stability of self-folded nanorackets and nanorings and demonstrate that such aggregated amyloid fibrils are stable in such states even when the fibril-fibril interaction is relatively weak, suggesting a strong propensity towards aggregation, given that the constituting amyloid fibril lengths exceed a critical fibril length-scale of several hundred nanometers. We further present a simple approach to directly determine the interfibril adhesion strength from geometric measures. In addition to providing insight into the physics of aggregation of amyloid fibrils our model enables the analysis of large-scale amyloid plaques and presents a new method for the estimation and engineering of the adhesive forces responsible of the self-assembly process of amyloid nanostructures, filling a 2 gap that previously existed between full atomistic simulations of primarily ultra-short fibrils and much larger micrometer-scale amyloid aggregates. Via direct simulation of large-scale amyloid plaques consisting of hundreds of fibrils we demonstrate that the fibril length has a profound impact on their structure and mechanical properties, where critical fibril length-scale derived from our analysis defines the structure of amyloid plaques. A multi-scale modeling approach as used here, bridging the scales from Angstroms to micrometers, opens a wide range of possible nanotechnology applications by presenting a holistic framework that balances mechanical properties of individual fibrils, hierarchical self-assembly, and the adhesive forces determining their stability to facilitate the design of de novo amyloid materials.

show abstract

Preparation and Characterization of Purified Amyloid Fibrils

Cited by 123 publications

References 10 publications

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Amyloid Fibril Formation by Lens Crystallin Proteins and Its Implications for Cataract Formation

Self-folding and aggregation of amyloid nanofibrils

Contact Info

Product

Resources

About