Instituto de Bioquímica Médica Leopoldo de Meis, Programa de Biologia Estrutural , Universidade Federal do Rio de Janeiro , Rio de Janeiro , RJ 21941-590 , Brazil.
Fiocruz Pernambuco , Instituto Aggeu Magalhães, Departamento de Microbiologia , Recife , PE 50740-465 , Brazil.
ACS Chem Neurosci. 2018 Nov 21;9(11):2807-2814. doi: 10.1021/acschemneuro.8b00222. Epub 2018 May 31.
Protein aggregation is a hallmark of several degenerative diseases, including Alzheimer's disease, Parkinson's disease and familial amyloidosis (Finnish type) (FAF). A method to isolate and detect amyloids is desired for the diagnosis of amyloid diseases. Here, we report the synthesis of pentameric thiophene amyloid ligand (p-FTAA) linked to agarose resin for selective purification of amyloid aggregates produced in vitro and in vivo. Using amyloid fibrils produced in vitro from α-synuclein, gelsolin, and Aβ and gelsolin amyloid aggregates extracted from tissue homogenates of a mouse model of FAF, we observed that p-FTAA resin was able to pull down amyloid aggregates. The functionalized resin was also able to pull down oligomers produced in vitro from the A30P variant of α-synuclein. The methodology described here can be useful for the diagnosis of amyloidogenic disease and also can be used to purify amyloid fibrils from biological samples, rendering the fibrils available for more accurate structural and biochemical characterization.
蛋白质聚集是几种退行性疾病的标志，包括阿尔茨海默病、帕金森病和家族性淀粉样变性（芬兰型）（FAF）。人们希望有一种分离和检测淀粉样蛋白的方法，用于淀粉样疾病的诊断。在这里，我们报告了五聚噻吩淀粉样配体（p-FTAA）与琼脂糖树脂的连接，用于选择性纯化体外和体内产生的淀粉样聚集物。使用来自α-突触核蛋白、凝胶蛋白和 Aβ的体外产生的淀粉样纤维以及从 FAF 小鼠模型组织匀浆中提取的凝胶蛋白淀粉样聚集物，我们观察到 p-FTAA 树脂能够沉淀淀粉样聚集物。功能化树脂还能够沉淀体外从α-突触核蛋白的 A30P 变体产生的寡聚物。这里描述的方法可用于淀粉样疾病的诊断，也可用于从生物样品中纯化淀粉样纤维，使纤维可用于更准确的结构和生化特性分析。
