Department of Pathology, New York University School of Medicine, New York, NY, USA.
Instituto de Salud Carlos III, Madrid, Spain.
Methods Mol Biol. 2022;2466:49-60. doi: 10.1007/978-1-0716-2176-9_4.
Affinity chromatography has, for many years, been at the research forefront as one of the simplest although highly versatile techniques capable of identifying biologically relevant protein-protein interactions. In the field of amyloid disorders, the use of ligands immobilized to a variety of affinity matrices was the method of choice to individualize proteins with affinity for soluble circulating forms of amyloid subunits. The methodology has also played an important role in the identification of proteins that interact with different amyloidogenic peptides and, as a result, are capable of modulating their physiological and pathological functions by altering solubility, aggregation propensity, and fibril formation proclivity. Along this line, classical studies conducted in the field of Alzheimer's disease (AD) identified clusterin as a major binding protein to both circulating soluble Aβ as well as to the brain deposited counterpart. The affinity chromatography-based approach employed herein, individualized clusterin as the major protein capable of binding the amyloid subunits associated with familial British and Danish dementias, two non-Aβ neurodegenerative conditions also exhibiting cerebral amyloid deposition and sharing striking similarities to AD. The data demonstrate that clusterin binding ability to amyloid molecules is not restricted to Aβ, suggesting a modulating effect on the aggregation/fibrillization propensity of the amyloidogenic peptides that is consistent with its known chaperone activity.
亲和层析多年来一直处于研究前沿，是一种最简单但功能非常强大的技术，能够识别与生物相关的蛋白质-蛋白质相互作用。在淀粉样蛋白紊乱领域，使用固定在各种亲和基质上的配体来个体化与可溶性循环淀粉样蛋白亚单位具有亲和力的蛋白质是首选方法。该方法在鉴定与不同淀粉样肽相互作用的蛋白质方面也发挥了重要作用，并且能够通过改变溶解度、聚集倾向和纤维形成倾向来调节其生理和病理功能。沿着这条线，在阿尔茨海默病（AD）领域进行的经典研究表明，簇蛋白是与循环可溶性 Aβ以及脑内沉积的对应物都有结合的主要结合蛋白。本文采用的基于亲和层析的方法将簇蛋白鉴定为与家族性英国和丹麦痴呆症相关的淀粉样蛋白亚单位具有结合能力的主要蛋白，这两种非 Aβ 神经退行性疾病也表现出脑淀粉样蛋白沉积，并与 AD 具有惊人的相似性。数据表明，簇蛋白对淀粉样蛋白分子的结合能力不仅限于 Aβ，这表明其对淀粉样肽聚集/纤维化倾向具有调节作用，这与其已知的伴侣活性一致。
