Nanomechanical Characterization of Apolipoprotein A-I Amyloid Fibrils

Amyloid fibrils represent a special type of protein aggregates that are currently receiving enormous attention due to their strong implication in molecular etiology of a wide range of human disorders. Amyloid fibrils represent highly ordered self-assemblies sharing a core cross-β-sheet structure. Such organization of the fibrils is responsible for amyloid insolubility and exceptional mechanical properties. The remarkable rigidity of the protein fibrillar aggregates is due to intra- and interstrand hydrogen bonds which stabilize the β-strand scaffold of amyloid fibrils. Increasing evidence indicates that physical properties of amyloid assemblies, especially their mechanical characteristics, play essential role in determining their cytotoxic action. This highlights the necessity of deciphering the correlation between the elastic properties of amyloid aggregates and their cytotoxicity. In the present paper we utilized the atomic force microscopy (AFM) to visualize and analyze the amyloid fibrils of G26R/W@8 mutant of N-terminal fragment of human apolipoprotein A-I (apoA-I). The examination of AFM images revealed the existence of two polymorphic forms of apoA-I fibrils – twisted ribbon and helical ribbon. The quantitative analysis of apoA-I elastic properties was performed within the framework of worm-like model of polymer chain using the Easyworm software. The Easyworm package analyzes the images of individual polymer chains obtained by the atomic force microscopy and allows calculation of the persistent length of a chain in three regimes depending on the ratio between the contour and persistent lengths of the polymer. The set of evaluated parameters included the Young’s modulus, persistent length, bending rigidity and the second moment of inertia. All parameters calculated for the helical ribbon conformation were higher than those of the twisted ribbon. These findings suggest that helical ribbon represents a more rigid and mechanically stable configuration. The results obtained may prove of importance for a deeper understanding the mechanics-driven pathological activities of amyloid fibrils.

PHYSICAL-CHEMICAL PROPERTIES OF CHONDROITIN SULFATE IN AGGRECAN

In current work theoretical study of physical-chemical properties of chondroitin sulfate in aggrecan within the self-consistent field theory was developed. As a model for aggrecan a model of cylindrical polymer brush was chosen. Aggrecan is considered as a system of n gaussian polymer chains consisting of N segments tethered to cylindrical surface of core protein. Basing on the results of previous work where using gaussian equivalent representation for functional integrals equation of state for chondroitin sulfate in aqueous solution was obtained and also osmotic pressure, dissociation degree, radius of gyration and renormalized persistent length were calculated with respect to monomers electrostatic interactions and effect of counterion condensation, one can obtain configuration integral for cylindrical polymer brush. Persistent length is introduced instead of Kuhn segment length which supposedly helps one to take into account rigidity of polymer chain determined by electrostatic repulsion of monomers (disaccharides groups). Expression for free energy of a polymer brush modeled as chondroitin sulfate chains tethered to a cylindrical surface was obtained. Dependence of chondroitin sulfate in aggrecan dissociation degree as a function of distance from the cylindrical surface is represented. For given system it varies insignificantly from the one for free chains in solution. Results for a polymer density as a function of distance from the cylindrical surface obtained within mean-field approximation are in qualitative agreement with those obtained using density functional method. However, calculations within a mean field framework are understable, thus it is necessary to use regularization methods.Forcitation:NogovitsynE.A., KalikinN.N., ZheleznyakN.I. Physical-chemical properties of chondroitin sulfate in aggrecan. Izv. Vyssh. Uchebn. Zaved. Khim. Khim. Tekhnol. 2018. V. 61. N 2. P. 35-39