An enzyme-linked immunosorbent assay to compare the affinity of chemical compounds for beta-amyloid peptide as a monomer

doi:10.1007/s00216-009-3420-6

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	An enzyme-linked immunosorbent assay to compare the affinity of chemical compounds for beta-amyloid peptide as a monomer
	Jiang, Chunyi 1; Feng, Yu 1; Huang, Xiaotong 3; Xu, Yechun2 ; Zhang, Yaping 4; Zhou, Naiming 4; Shen, Xu1 ; Chen, Kaixian2 ; Jiang, Hualiang2 ; Liu, Dongxiang1
刊名	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
	2010-03
卷号	396 期号:5 页码:1745-1754
关键词	Enzyme-linked immunosorbent assay beta-Amyloid peptide Binding affinity Inhibitor Bcl-x(L)
ISSN号	1618-2642
DOI	10.1007/s00216-009-3420-6
文献子类	Article
英文摘要	A beta(1-42) is the proteolytic cleavage product of cleavage of the amyloid precursor protein by beta- and gamma- secretases. The aggregation of A beta(1-42) plays a causative role in the development of Alzheimer's disease. To lock A beta(1-42) in a homogenous state, we embedded the A beta(1-42) sequence in an unstructured region of Bcl-x(L). Both the N-terminus and the C-terminus of A beta(1-42) were constrained in the disordered region, whereas the conjunction did not introduce any folding to A beta(1-42) but maintained the sequence as a monomer in solution. With Bcl-x(L)-A beta(42), we developed an enzyme-linked immunosorbent assay to compare the affinity of compounds for monomeric A beta(1-42). Bcl-x(L)-A beta(42) was coated on a microplate and this was followed by incubation with different concentrations of compounds. Compounds binding to Leu17-Val24 of A beta(1-42) inhibited the interaction between Bcl-x(L)-A beta(42) and antibody 4G8. The method can not only reproduce the activities of the reported A beta(1-42) inhibitors such as dopamine, tannin, and morin but can also differentiate decoy compounds that do not bind to A beta(1-42). Remarkably, using this method, we discovered a new inhibitor that binds to monomeric A beta(1-42) and inhibits A beta(1-42) fibril formation. As the structure of Bcl-x(L)-A beta(42) monomer is stable in solution, the assay could be adapted for high-throughput screening with a series of antibodies that bind the different epitopes of A beta(1-42). In addition, the monomeric form of the A beta(1-42) sequence in Bcl-x(L)-A beta(42) would also facilitate the identification of A beta(1-42) binding partners by coimmunoprecipitation, cocrystallization, surface plasmon resonance technology, or the assay as described here.
资助项目	CAS Introducing Outstanding Overseas Scientists Project[00000000] ; NSFC Innovation Program for Research Group[20721003] ; National Science & Technology Major Project[2009ZX09301-001]
WOS关键词	ALZHEIMERS-DISEASE ; IN-VITRO ; FIBRILS ; AGGREGATION ; PROTEIN ; NMR ; FIBRILLOGENESIS ; CYCLODEXTRIN ; INHIBITOR ; MECHANISM
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	SPRINGER HEIDELBERG
WOS记录号	WOS:000274742900016
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/278955]
专题	药物发现与设计中心药理学第三研究室
通讯作者	Liu, Dongxiang
作者单位	1.Chinese Acad Sci, Dept Mol Pharmacol, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China; 2.Chinese Acad Sci, Ctr Drug Design & Discovery, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China; 3.Singapore Polytech, Sch Chem & Life Sci, Singapore 139651, Singapore; 4.Zhejiang Univ, Inst Biochem, Coll Life Sci, Hangzhou 310058, Zhejiang, Peoples R China
推荐引用方式 GB/T 7714	Jiang, Chunyi,Feng, Yu,Huang, Xiaotong,et al. An enzyme-linked immunosorbent assay to compare the affinity of chemical compounds for beta-amyloid peptide as a monomer[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2010,396(5):1745-1754.
APA	Jiang, Chunyi.,Feng, Yu.,Huang, Xiaotong.,Xu, Yechun.,Zhang, Yaping.,...&Liu, Dongxiang.(2010).An enzyme-linked immunosorbent assay to compare the affinity of chemical compounds for beta-amyloid peptide as a monomer.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,396(5),1745-1754.
MLA	Jiang, Chunyi,et al."An enzyme-linked immunosorbent assay to compare the affinity of chemical compounds for beta-amyloid peptide as a monomer".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 396.5(2010):1745-1754.