一种用于组蛋白乙酰转移酶活性的快速灵敏检测方法。
A rapid and sensitive assay for histone acetyl-transferase activity.
作者信息
Ait-Si-Ali S, Ramirez S, Robin P, Trouche D, Harel-Bellan A
机构信息
Laboratoire Oncogénèse, Différenciation et Transduction du Signal, CNRS UPR 9079, IFC-01, 7 rue Guy Moquet, 94801 Villejuif, France.
出版信息
Nucleic Acids Res. 1998 Aug 15;26(16):3869-70. doi: 10.1093/nar/26.16.3869.
Abstract
Histone acetyl-transferases (HATs) seem to be key elements in the regulation of transcription. We have designed an enzymatic assay to quantify HAT enzymatic activity. In this assay, the substrate is a peptide corresponding to the 24 first amino acids of histone H4 which is coupled to biotin. After acetylation using [14C]acetyl-CoA, the peptide is purified on streptavidin beads and the associated radioactivity is measured. This assay is sensitive, rapid and convenient.
摘要
组蛋白乙酰转移酶（HATs）似乎是转录调控中的关键因素。我们设计了一种酶活性测定方法来定量HAT的酶活性。在该测定中，底物是与生物素偶联的对应于组蛋白H4前24个氨基酸的肽。使用[14C]乙酰辅酶A进行乙酰化后，该肽在链霉亲和素珠上纯化，并测量相关的放射性。该测定方法灵敏、快速且方便。
相似文献
1
A rapid and sensitive assay for histone acetyl-transferase activity.一种用于组蛋白乙酰转移酶活性的快速灵敏检测方法。
Nucleic Acids Res. 1998 Aug 15;26(16):3869-70. doi: 10.1093/nar/26.16.3869.
2
Application of a fluorescent histone acetyltransferase assay to probe the substrate specificity of the human p300/CBP-associated factor.应用荧光组蛋白乙酰转移酶测定法探究人类p300/CBP相关因子的底物特异性。
Anal Biochem. 2000 Dec 15;287(2):319-28. doi: 10.1006/abio.2000.4855.
3
Functional analysis of the p300 acetyltransferase domain: the PHD finger of p300 but not of CBP is dispensable for enzymatic activity.p300 乙酰转移酶结构域的功能分析：p300 的 PHD 结构域而非 CBP 的 PHD 结构域对酶活性而言并非必需。
Nucleic Acids Res. 2001 Nov 1;29(21):4462-71. doi: 10.1093/nar/29.21.4462.
4
Identification and analysis of yeast nucleosomal histone acetyltransferase complexes.酵母核小体组蛋白乙酰转移酶复合物的鉴定与分析。
Methods. 1998 Aug;15(4):315-21. doi: 10.1006/meth.1998.0635.
5
Site specificity analysis of Piccolo NuA4-mediated acetylation for different histone complexes.小皮琴（Piccolo）NuA4介导的不同组蛋白复合物乙酰化的位点特异性分析。
Biochem J. 2015 Dec 1;472(2):239-48. doi: 10.1042/BJ20150654. Epub 2015 Sep 29.
6
Site specificity of pea histone acetyltransferase B in vitro.豌豆组蛋白乙酰转移酶B的体外位点特异性
J Biol Chem. 1993 Jun 25;268(18):13248-52.
7
A continuous, nonradioactive assay for histone acetyltransferases.一种用于组蛋白乙酰转移酶的连续非放射性检测方法。
Anal Biochem. 2000 May 1;280(2):308-14. doi: 10.1006/abio.2000.4546.
8
Purified histone acetyltransferase complexes stimulate HIV-1 transcription from preassembled nucleosomal arrays.纯化的组蛋白乙酰转移酶复合物可刺激HIV-1从预先组装的核小体阵列进行转录。
Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):12924-9. doi: 10.1073/pnas.95.22.12924.
9
Histone modification patterns during gene activation.基因激活过程中的组蛋白修饰模式。
Methods Enzymol. 2004;377:130-53. doi: 10.1016/S0076-6879(03)77007-4.
10
Properties of Nat4, an N(alpha)-acetyltransferase of Saccharomyces cerevisiae that modifies N termini of histones H2A and H4.酿酒酵母N（α）-乙酰转移酶Nat4的特性，该酶可修饰组蛋白H2A和H4的N端。
Mol Cell Biol. 2009 Jun;29(11):2913-24. doi: 10.1128/MCB.00147-08. Epub 2009 Mar 30.
引用本文的文献
1
Advances in direct detection of lysine methylation and acetylation by nuclear magnetic resonance using C-enriched cofactors.利用富含碳的辅助因子通过核磁共振直接检测赖氨酸甲基化和乙酰化的进展。
Methods. 2023 Oct;218:72-83. doi: 10.1016/j.ymeth.2023.07.010. Epub 2023 Jul 29.
2
A direct nuclear magnetic resonance method to investigate lysine acetylation of intrinsically disordered proteins.一种用于研究内在无序蛋白质赖氨酸乙酰化的直接核磁共振方法。
Front Mol Biosci. 2023 Jan 6;9:1074743. doi: 10.3389/fmolb.2022.1074743. eCollection 2022.
3
Ultrasensitive electrochemiluminescence immunosensor for the transcriptional co-activator p300 by using a graphene oxide monolayer and tetrahedral DNA-mediated signal amplification.基于氧化石墨烯单层和四面体 DNA 介导的信号放大的转录共激活因子 p300 的超灵敏电化学发光免疫传感器
Mikrochim Acta. 2019 May 2;186(5):325. doi: 10.1007/s00604-019-3435-4.
4
Acetylation- and Methylation-Related Epigenetic Proteins in the Context of Their Targets.与其靶点相关的乙酰化和甲基化相关表观遗传蛋白
Genes (Basel). 2017 Aug 7;8(8):196. doi: 10.3390/genes8080196.
5
Epigenetic assays for chemical biology and drug discovery.用于化学生物学和药物发现的表观遗传学检测
Clin Epigenetics. 2017 Apr 21;9:41. doi: 10.1186/s13148-017-0342-6. eCollection 2017.
6
Effective Quenchers Are Required to Eliminate the Interference of Substrate: Cofactor Binding in the HAT Scintillation Proximity Assay.需要有效的淬灭剂来消除底物干扰：HAT闪烁邻近分析中的辅因子结合。
Assay Drug Dev Technol. 2015 May;13(4):210-20. doi: 10.1089/adt.2015.636. Epub 2015 May 28.
7
An inactive geminin mutant that binds cdt1.一种失活的 geminin 突变体，能与 cdt1 结合。
Genes (Basel). 2015 May 15;6(2):252-66. doi: 10.3390/genes6020252.
8
PAINS in the assay: chemical mechanisms of assay interference and promiscuous enzymatic inhibition observed during a sulfhydryl-scavenging HTS.检测中的问题：在巯基清除高通量筛选过程中观察到的检测干扰和混杂酶抑制的化学机制。
J Med Chem. 2015 Mar 12;58(5):2091-113. doi: 10.1021/jm5019093. Epub 2015 Feb 21.
9
A cell-free fluorometric high-throughput screen for inhibitors of Rtt109-catalyzed histone acetylation.一种无细胞荧光高通量筛选 Rtt109 催化组蛋白乙酰化抑制剂的方法。
PLoS One. 2013 Nov 18;8(11):e78877. doi: 10.1371/journal.pone.0078877. eCollection 2013.
10
Comparative studies of thiol-sensitive fluorogenic probes for HAT assays.硫醇敏感性荧光探针用于 HAT 测定的比较研究。
Anal Bioanal Chem. 2013 Feb;405(4):1361-71. doi: 10.1007/s00216-012-6522-5. Epub 2012 Nov 9.
本文引用的文献
1
Rb interacts with histone deacetylase to repress transcription.Rb与组蛋白去乙酰化酶相互作用以抑制转录。
Cell. 1998 Feb 20;92(4):463-73. doi: 10.1016/s0092-8674(00)80940-x.
2
Retinoblastoma protein represses transcription by recruiting a histone deacetylase.视网膜母细胞瘤蛋白通过招募组蛋白去乙酰化酶来抑制转录。
Nature. 1998 Feb 5;391(6667):601-5. doi: 10.1038/35410.
3
Retinoblastoma protein recruits histone deacetylase to repress transcription.视网膜母细胞瘤蛋白招募组蛋白去乙酰化酶以抑制转录。
Nature. 1998 Feb 5;391(6667):597-601. doi: 10.1038/35404.
4
Histone deacetylase activity is required for full transcriptional repression by mSin3A.组蛋白去乙酰化酶活性是mSin3A实现完全转录抑制所必需的。
Cell. 1997 May 2;89(3):341-7. doi: 10.1016/s0092-8674(00)80214-7.
5
The CBP co-activator is a histone acetyltransferase.CBP共激活因子是一种组蛋白乙酰转移酶。
Nature. 1996;384(6610):641-3. doi: 10.1038/384641a0.
6
The transcriptional coactivators p300 and CBP are histone acetyltransferases.转录共激活因子p300和CBP是组蛋白乙酰转移酶。
Cell. 1996 Nov 29;87(5):953-9. doi: 10.1016/s0092-8674(00)82001-2.
7
Histone acetylation and chromatin assembly: a single escort, multiple dances?组蛋白乙酰化与染色质组装：一位单一的护送者，多场舞蹈？
Cell. 1996 Oct 4;87(1):5-8. doi: 10.1016/s0092-8674(00)81316-1.
8
A mammalian histone deacetylase related to the yeast transcriptional regulator Rpd3p.一种与酵母转录调节因子Rpd3p相关的哺乳动物组蛋白脱乙酰基酶。
Science. 1996 Apr 19;272(5260):408-11. doi: 10.1126/science.272.5260.408.
9
Tetrahymena histone acetyltransferase A: a homolog to yeast Gcn5p linking histone acetylation to gene activation.嗜热四膜虫组蛋白乙酰转移酶A：一种与酵母Gcn5p同源的蛋白，将组蛋白乙酰化与基因激活联系起来。
Cell. 1996 Mar 22;84(6):843-51. doi: 10.1016/s0092-8674(00)81063-6.
10
Targeting chromatin disruption: Transcription regulators that acetylate histones.靶向染色质破坏：使组蛋白乙酰化的转录调节因子。
Cell. 1996 Mar 22;84(6):817-9. doi: 10.1016/s0092-8674(00)81059-4.
Zotero 插件