ArticleAnalytical chemistry2011
Characterizing DNA methyltransferases with an ultrasensitive luciferase-linked continuous assay.
Article in Analytical chemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 42 citations in OpenAlex.
- Chemical probes and methods for the study of protein arginine methylation.RSC chemical biology · 2023Review
- Locally correlated kinetics of post-replication DNA methylation reveals processivity and region specificity in DNA methylation maintenance.Journal of the Royal Society, Interface · 2022Article
- Chemical biology and medicinal chemistry of RNA methyltransferases.Nucleic acids research · 2022Article
- Synthesis and Characterization of Transition-State Analogue Inhibitors against Human DNA Methyltransferase 1.Journal of medicinal chemistry · 2022Article
- The acute myeloid leukemia variant DNMT3A Arg882His is a DNMT3B-like enzyme.Nucleic acids research · 2020Article
- Identification of the mMolecular cell · 2019Article
- Lysine methylation signaling of non-histone proteins in the nucleus.Cellular and molecular life sciences : CMLS · 2019Review
- Direct Detection of Products from S-Adenosylmethionine-Dependent Enzymes Using a Competitive Fluorescence Polarization Assay.Analytical chemistry · 2018Article
- Hydrogen Peroxide-Based Fluorometric Assay for Real-Time Monitoring of SAM-Dependent Methyltransferases.Frontiers in bioengineering and biotechnology · 2018Article
- A simplified characterization ofChemical science · 2017Article
- In Utero Exposure to a High-Fat Diet Programs Hepatic Hypermethylation and Gene Dysregulation and Development of Metabolic Syndrome in Male Mice.Endocrinology · 2017Article
- Resveratrol-salicylate derivatives as selective DNMT3 inhibitors and anticancer agents.Journal of enzyme inhibition and medicinal chemistry · 2016Article
- Human DNMT1 transition state structure.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Small Molecule Inhibitors of Protein Arginine Methyltransferases.Expert opinion on investigational drugs · 2016Review
- A Comparative Analysis of Acyl-Homoserine Lactone Synthase Assays.Chembiochem : a European journal of chemical biology · 2015Article
- Histone H2A and H4 N-terminal tails are positioned by the MEP50 WD repeat protein for efficient methylation by the PRMT5 arginine methyltransferase.The Journal of biological chemistry · 2015Article
- A sensitive luminescent assay for the histone methyltransferase NSD1 and other SAM-dependent enzymes.Assay and drug development technologies · 2014Article
- Bioluminescence: a versatile technique for imaging cellular and molecular features.MedChemComm · 2014Article
- Development of a universal radioactive DNA methyltransferase inhibition test for high-throughput screening and mechanistic studies.Nucleic acids research · 2013Article
- Luciferase-based assay for adenosine: application to S-adenosyl-L-homocysteine hydrolase.Analytical chemistry · 2012Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
DNA (cytosine-5)-methyltransferases (DNMTs) catalyze the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to the 5-position of cytosine residues and thereby silence transcription of regulated genes. DNMTs are important epigenetic targets. However, isolated DNMTs are weak catalysts and are difficult to assay. We report an ultrasensitive luciferase-linked continuous assay that converts the S-adenosyl-L-homocysteine product of DNA methylation to a quantifiable luminescent signal. Results with this assay are compared with the commonly used DNA labeling from [methyl-(3)H]AdoMet. A 5'-methylthioadenosine-adenosylhomocysteine nucleosidase is used to hydrolyze AdoHcy to adenine. Adenine phosphoribosyl transferase converts adenine to AMP and pyruvate orthophosphate dikinase converts AMP to ATP. Firefly luciferase gives a stable luminescent signal that results from continuous AMP recycling to ATP. This assay exhibits a broad dynamic range (0.1-1000 pmol of AdoHcy). The rapid response time permits continuous assays of DNA methylation detected by light output. The assay is suitable for high-throughput screening of chemical libraries for DNMT inhibition activity. The kinetic properties of human and bacterial CpG methyltransferases are characterized using this assay. Human catalytic domain DNMT3b activation by DNMT3L is shown to involve two distinct kinetic states that alter k(cat) but not K(m) for AdoMet. The assay is shown to be robust in the presence of high concentrations of the pyrimidine analogues 5-azacytidine and 5-azacytosine.
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