南方科技大学知识苑(SUSTech KC): FRET-Based Method for Direct, Real-Time Measurement of DNA Methyltransferase Activity

题名	FRET-Based Method for Direct, Real-Time Measurement of DNA Methyltransferase Activity
作者	Long, Yi1,2 ; Ubych, Krystian 1; Jagu, Elodie 1,3; Neely, Robert K.1
通讯作者	Neely, Robert K.
发表日期	2021-01-20
DOI	10.1021/acs.bioconjchem.0c00612
发表期刊	BIOCONJUGATE CHEMISTRY 影响因子和分区
ISSN	1043-1802
EISSN	1520-4812
卷号	32 期号:1页码:192-198
摘要	DNA methyltransferase activity is associated with a host of diseases, including cancers, where global hypomethylation of the genome, as well as marked changes in local DNA methylation patterns, can be both diagnostic and prognostic for the disease. Despite this, we currently lack a method for directly measuring the activity of the DNA methyltransferases, which would support the development of DNA methyltransferase-targeted therapies. Here, we demonstrate an assay for the direct measurement of methyltransferase activity, in real time. We employ a fluorescent methyltransferase cofactor analogue, which when bound by the enzyme to a labeled target DNA sequence results in fluorescence resonance energy transfer (FRET) between the donor dye (DNA) and the acceptor dye (cofactor). We demonstrate that the method can be used to monitor the activity of DNA MTases in real time and can be applied to screen inhibitors of the DNA methyltransferases. We show this in both bulk phase and single molecule imaging experiments, highlighting the potential application of the assay in screening and biophysical studies of methyltransferase function.
相关链接	[来源记录]
收录类别	SCI ; EI
语种	英语
学校署名	其他
资助项目	Engineering and Physical Sciences Research Council (EPSRC)[EP/N020901/1]
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary ; Chemistry, Organic
WOS记录号	WOS:000612551500020
出版者	AMER CHEMICAL SOC
EI入藏号	20220411530246
ESI学科分类	CHEMISTRY
来源库	Web of Science
引用统计	被引频次[WOS]：9
成果类型	期刊论文
条目标识符	http://sustech.caswiz.com/handle/2SGJ60CL/210878
专题	南方科技大学医院
作者单位	1.Univ Birmingham, Sch Chem, Birmingham B15 2TT, W Midlands, England 2.Southern Univ Sci & Technol Hosp, Med Res Ctr, Shenzhen 518055, Guangdong, Peoples R China 3.Univ Clermont Auvergne, CNRS, SIGMA Clermont, ICCF, F-63178 Aubiere, France
第一作者单位	南方科技大学医院
推荐引用方式 GB/T 7714	Long, Yi,Ubych, Krystian,Jagu, Elodie,et al. FRET-Based Method for Direct, Real-Time Measurement of DNA Methyltransferase Activity[J]. BIOCONJUGATE CHEMISTRY,2021,32(1):192-198.
APA	Long, Yi,Ubych, Krystian,Jagu, Elodie,&Neely, Robert K..(2021).FRET-Based Method for Direct, Real-Time Measurement of DNA Methyltransferase Activity.BIOCONJUGATE CHEMISTRY,32(1),192-198.
MLA	Long, Yi,et al."FRET-Based Method for Direct, Real-Time Measurement of DNA Methyltransferase Activity".BIOCONJUGATE CHEMISTRY 32.1(2021):192-198.