A fluorometric assay of SIRT1 deacetylation activity through quantification of nicotinamide adenine dinucleotide
文献类型：期刊论文
| 作者 | Feng, Yu1; Wu, Jiahui1; Chen, Lei1; Luo, Chen2; Shen, Xu1 ; Chen, Kaixian2; Jiang, Hualiang2; Liu, Dongxiang1
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| 刊名 | ANALYTICAL BIOCHEMISTRY
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| 出版日期 | 2009-12-15 |
| 卷号 | 395期号:2页码:205-210 |
| 关键词 | Sirtuins NAD(+) Deacetylation Fluorescence Cyclized alpha-adduct |
| ISSN号 | 0003-2697 |
| DOI | 10.1016/j.ab.2009.08.011 |
| 文献子类 | Article |
| 英文摘要 | Sirtuins are nicotinamide adenine dinucleotide (NAD(+))-dependent deacetylases that catalyze the deacetylation of proteins Such as histones and p53. A sensitive and convenient fluorometric assay for evaluating the SIRT1 enzymatic activity was developed here. Specifically, the remaining NAD(+) after the deacetylation was determined by converting NAD(+) to a highly fluorescent cyclized alpha-adduct compound. By this assay, we found that nicotinamide, Cu2+, and Zn2+ antagonize the activity of SIRT1. Resveratrol stimulates the enzymatic activity specifically with 7-amino-4-methylcoumarin (AMC)-labeled acetylated peptide, Epigallocatechin galate (EGCG) inhibits SIRT1 activity with both AMC-labeled and unlabeled peptide. However, a combination of vitamin C with EGCG can reverse the inhibition of EGCG with the unlabeled peptide or stimulate the deacetylation of AMC-labeled peptide by SIRT1. The assay does not require any isotopic material and thus is biologically safe. It can be adapted to a 96-well microplate for high-throughput Screening. Notably, the acetylated peptides with or without fluorescent labels may be used in the assay, which facilitates the Substrate specificity study of SIRT1 activators or inhibitors in vitro. (C) 2009 Elsevier Inc. All rights reserved. |
| WOS关键词 | SMALL-MOLECULE ACTIVATORS ; NAD(+)-DEPENDENT DEACETYLASES ; HISTONE/PROTEIN DEACETYLASES ; PROTEIN DEACETYLASES ; LIFE-SPAN ; FAMILY ; SUBSTRATE ; RESVERATROL ; INHIBITOR ; MECHANISM |
| 资助项目 | CAS Introducing Outstanding Oversea Scientists Project[00000000] ; National Science Fund for Creative Research Group[20721003] ; Science and Technology Commission of Shanghai Municipality[08JC1422100] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000271173600011 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/279062]  |
| 专题 | 药理学第三研究室 药物发现与设计中心 |
| 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, Shanghai Inst Mat Med, Ctr Drug Design & Discovery, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Feng, Yu,Wu, Jiahui,Chen, Lei,et al. A fluorometric assay of SIRT1 deacetylation activity through quantification of nicotinamide adenine dinucleotide[J]. ANALYTICAL BIOCHEMISTRY,2009,395(2):205-210. |
| APA | Feng, Yu.,Wu, Jiahui.,Chen, Lei.,Luo, Chen.,Shen, Xu.,...&Liu, Dongxiang.(2009).A fluorometric assay of SIRT1 deacetylation activity through quantification of nicotinamide adenine dinucleotide.ANALYTICAL BIOCHEMISTRY,395(2),205-210. |
| MLA | Feng, Yu,et al."A fluorometric assay of SIRT1 deacetylation activity through quantification of nicotinamide adenine dinucleotide".ANALYTICAL BIOCHEMISTRY 395.2(2009):205-210. |
入库方式： OAI收割
来源：上海药物研究所
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