Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model.
An initial step in amyloid-β (Aβ) production includes amyloid precursor protein (APP) cleavage via β-Site amyloid precursor protein cleaving enzyme 1 (BACE1). Increased levels of brain Aβ have been implicated in the pathogenesis of Alzheimer's disease (AD). Thus, β-secretase represents a primar...
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doaj-4097e5d236f743459d33adea7ce995562020-11-25T02:13:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011010e014073310.1371/journal.pone.0140733Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model.Huiyu LiangYusheng ShiZhewen KouYonghua PengWenjun ChenXiaowen LiShuji LiYing WangFang WangXingmei ZhangAn initial step in amyloid-β (Aβ) production includes amyloid precursor protein (APP) cleavage via β-Site amyloid precursor protein cleaving enzyme 1 (BACE1). Increased levels of brain Aβ have been implicated in the pathogenesis of Alzheimer's disease (AD). Thus, β-secretase represents a primary target for inhibitor drug development in AD. In this study, aptamers were obtained from combinatorial oligonucleotide libraries using a technology referred to as systematic evolution of ligands by exponential enrichment (SELEX). A purified human BACE1 extracellular domain was used as a target to conduct an in vitro selection process using SELEX. Two DNA aptamers were capable of binding to BACE1 with high affinity and good specificity, with Kd values in the nanomolar range. We subsequently confirmed that one aptamer, A1, exhibited a distinct inhibitory effect on BACE1 activity in an AD cell model. We detected the effects of M17-APPsw cells that stably expressed Swedish mutant APP after aptamer A1 treatment. Aβ40 and Aβ42 concentrations secreted by M17-APPsw cells decreased intracellularly and in culture media. Furthermore, Western blot analysis indicated that sAPPβ expression significantly decreased in the A1 treated versus control groups. These findings support the preliminary feasibility of an aptamer evolved from a SELEX strategy to function as a potential BACE1 inhibitor. To our knowledge, this is the first study to acquire a DNA aptamer that exhibited binding specificity to BACE1 and inhibited its activity.http://europepmc.org/articles/PMC4608735?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Huiyu Liang Yusheng Shi Zhewen Kou Yonghua Peng Wenjun Chen Xiaowen Li Shuji Li Ying Wang Fang Wang Xingmei Zhang |
spellingShingle |
Huiyu Liang Yusheng Shi Zhewen Kou Yonghua Peng Wenjun Chen Xiaowen Li Shuji Li Ying Wang Fang Wang Xingmei Zhang Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. PLoS ONE |
author_facet |
Huiyu Liang Yusheng Shi Zhewen Kou Yonghua Peng Wenjun Chen Xiaowen Li Shuji Li Ying Wang Fang Wang Xingmei Zhang |
author_sort |
Huiyu Liang |
title |
Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. |
title_short |
Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. |
title_full |
Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. |
title_fullStr |
Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. |
title_full_unstemmed |
Inhibition of BACE1 Activity by a DNA Aptamer in an Alzheimer's Disease Cell Model. |
title_sort |
inhibition of bace1 activity by a dna aptamer in an alzheimer's disease cell model. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2015-01-01 |
description |
An initial step in amyloid-β (Aβ) production includes amyloid precursor protein (APP) cleavage via β-Site amyloid precursor protein cleaving enzyme 1 (BACE1). Increased levels of brain Aβ have been implicated in the pathogenesis of Alzheimer's disease (AD). Thus, β-secretase represents a primary target for inhibitor drug development in AD. In this study, aptamers were obtained from combinatorial oligonucleotide libraries using a technology referred to as systematic evolution of ligands by exponential enrichment (SELEX). A purified human BACE1 extracellular domain was used as a target to conduct an in vitro selection process using SELEX. Two DNA aptamers were capable of binding to BACE1 with high affinity and good specificity, with Kd values in the nanomolar range. We subsequently confirmed that one aptamer, A1, exhibited a distinct inhibitory effect on BACE1 activity in an AD cell model. We detected the effects of M17-APPsw cells that stably expressed Swedish mutant APP after aptamer A1 treatment. Aβ40 and Aβ42 concentrations secreted by M17-APPsw cells decreased intracellularly and in culture media. Furthermore, Western blot analysis indicated that sAPPβ expression significantly decreased in the A1 treated versus control groups. These findings support the preliminary feasibility of an aptamer evolved from a SELEX strategy to function as a potential BACE1 inhibitor. To our knowledge, this is the first study to acquire a DNA aptamer that exhibited binding specificity to BACE1 and inhibited its activity. |
url |
http://europepmc.org/articles/PMC4608735?pdf=render |
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