High-throughput screening-compatible assays of As(III) S-adenosylmethionine methyltransferase activity

doi:10.1016/j.ab.2015.04.011

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	High-throughput screening-compatible assays of As(III) S-adenosylmethionine methyltransferase activity
	Dong, Hui ; Xu, Wenzhong 1; Pillai, Jitesh K.; Packianathan, Charles ; Rosen, Barry P.
刊名	ANALYTICAL BIOCHEMISTRY
	2015
卷号	480 页码:67-73
关键词	Arsenic Methylation AS3MT High-throughput screens
ISSN号	0003-2697
DOI	10.1016/j.ab.2015.04.011
文献子类	Article
英文摘要	Arsenic is a naturally existing toxin and carcinogen. As(III) S-adenosylmethionine methyltransferases (AS3MT in mammals and ArsM in microbes) methylate As(III) three times in consecutive steps and play a central role in arsenic metabolism from bacteria to humans. Current assays for arsenic methylation are slow, laborious, and expensive. Here we report the development of two in vitro assays for AS3MT activity that are rapid, sensitive, convenient, and relatively inexpensive and can be adapted for high-throughput assays. The first assay measures As(III) binding by the quenching of the protein fluorescence of a single-tryptophan derivative of an AS3MT ortholog. The second assay utilizes time-resolved fluorescence resonance energy transfer to directly measure the conversion of the AS3MT substrate, S-adenosylmethionine, to S-adenosylhomocysteine catalyzed by AS3MT. These two assays are complementary, one measuring substrate binding and the other catalysis, making them useful tools for functional studies and future development of drugs to prevent arsenic-related diseases. (C) 2015 Elsevier Inc. All rights reserved.
学科主题	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
电子版国际标准刊号	1096-0309
出版地	SAN DIEGO
WOS关键词	TOXICITY ; BIOTRANSFORMATION ; BIOMETHYLATION ; IDENTIFICATION ; METHYLATION ; SINEFUNGIN ; MECHANISM ; EXPOSURE ; DISEASE ; TARGET
WOS研究方向	Science Citation Index Expanded (SCI-EXPANDED)
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000354912300012
资助机构	State of Florida Department of Health [4BF01] ; NIH from the National Institute of Environmental Health Sciences [R01 ES023779] ; Chinese Academy of Sciences ; NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES [R01ES023779] Funding Source: NIH RePORTER
内容类型	期刊论文
源URL	[http://ir.ibcas.ac.cn/handle/2S10CLM1/25873]
专题	中科院北方资源植物重点实验室
作者单位	1.[Dong, Hui; Pillai, Jitesh K.; Packianathan, Charles; Rosen, Barry P.] Florida Int Univ, Herbert Wertheim Coll Med, Dept Cellular Biol & Pharmacol, Miami, FL 33199 USA 2.Chinese Acad Sci, Key Lab Plant Resources, Inst Bot, Beijing 100093, Peoples R China
推荐引用方式 GB/T 7714	Dong, Hui,Xu, Wenzhong,Pillai, Jitesh K.,et al. High-throughput screening-compatible assays of As(III) S-adenosylmethionine methyltransferase activity[J]. ANALYTICAL BIOCHEMISTRY,2015,480:67-73.
APA	Dong, Hui,Xu, Wenzhong,Pillai, Jitesh K.,Packianathan, Charles,&Rosen, Barry P..(2015).High-throughput screening-compatible assays of As(III) S-adenosylmethionine methyltransferase activity.ANALYTICAL BIOCHEMISTRY,480,67-73.
MLA	Dong, Hui,et al."High-throughput screening-compatible assays of As(III) S-adenosylmethionine methyltransferase activity".ANALYTICAL BIOCHEMISTRY 480(2015):67-73.