Authentic hSAA related with AA amyloidosis: New method of purification, folding and amyloid polymorphism

IF 3.3 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Natalya Katina , Victor Marchenkov , Yulia Lapteva , Vitalii Balobanov , Nelly Ilyina , Natalya Ryabova , Stanislav Evdokimov , Mariya Suvorina , Alexey Surin , Anatoly Glukhov
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引用次数: 0

Abstract

The secondary amyloidosis of humans is caused by the formation of hSAA fibrils in different organs and tissues. Until now hSAA was thought to have low amyloidogenicity in vitro and the majority of SAA aggregation experiments were done using murine protein or hSAA non-pathogenic isoforms. In this work a novel purification method for recombinant hSAA was introduced, enabling to obtain monomeric protein capable of amyloid aggregation under physiological conditions. The stability and amyloid aggregation of hSAA have been examined using a wide range of biophysical methods. It was shown that the unfolding of monomeric protein occurs through the formation of molten globule-like intermediate state. Polymorphism of hSAA amyloids was discovered to depend on the solution pH. At pH 8.5, rapid protein aggregation occurs, which leads to the formation of twisted short fibrils. Even a slight decrease of the pH to 7.8 results in delayed aggregation with the formation of long straight amyloids composed of laterally associated protofilaments. Limited proteolysis experiments have shown that full-length hSAA is involved in the formation of intermolecular interactions in both amyloid polymorphs. The results obtained, and the experimental approach used in this study can serve as a basis for further research on the mechanism of authentic hSAA amyloid formation.

与 AA 淀粉样变性有关的真品 hSAA:纯化、折叠和淀粉样蛋白多态性的新方法
人类的继发性淀粉样变性是由 hSAA 纤维在不同器官和组织中的形成引起的。迄今为止,人们一直认为 hSAA 在体外的淀粉样蛋白致性较低,而且大多数 SAA 聚集实验都是使用鼠蛋白或 hSAA 非致病异构体进行的。在这项工作中,引入了一种新的重组 hSAA 纯化方法,从而获得了能够在生理条件下发生淀粉样聚集的单体蛋白。使用多种生物物理方法对 hSAA 的稳定性和淀粉样聚集进行了研究。研究表明,单体蛋白的解折是通过形成熔融球状中间状态来实现的。研究发现,hSAA淀粉样蛋白的多态性取决于溶液的pH值。在 pH 值为 8.5 时,蛋白质迅速聚集,形成扭曲的短纤维。即使 pH 值稍微降低到 7.8,也会导致延迟聚集,形成由横向关联的原丝组成的长直淀粉样。有限的蛋白水解实验表明,全长 hSAA 参与了两种淀粉样多态体中分子间相互作用的形成。本研究中获得的结果和使用的实验方法可作为进一步研究真正的 hSAA 淀粉样蛋白形成机制的基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biophysical chemistry
Biophysical chemistry 生物-生化与分子生物学
CiteScore
6.10
自引率
10.50%
发文量
121
审稿时长
20 days
期刊介绍: Biophysical Chemistry publishes original work and reviews in the areas of chemistry and physics directly impacting biological phenomena. Quantitative analysis of the properties of biological macromolecules, biologically active molecules, macromolecular assemblies and cell components in terms of kinetics, thermodynamics, spatio-temporal organization, NMR and X-ray structural biology, as well as single-molecule detection represent a major focus of the journal. Theoretical and computational treatments of biomacromolecular systems, macromolecular interactions, regulatory control and systems biology are also of interest to the journal.
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