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HotSpot: A Novel Methyltransferase Assay Platform for Drug Screening and Discovery
Author(s) -
Horiuchi Kurumi Y,
Eason Mia M,
Ma Haiching
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb61
Subject(s) - methyltransferase , epigenetics , drug discovery , histone , profiling (computer programming) , computational biology , chemistry , methylation , biology , dna , biochemistry , computer science , gene , operating system
Epigenetic regulation has been studied extensively in the past few years, and many studies indicate that it is strictly involved in tumor establishment and progression. Namely, Histone methyltransferases (HMTs) and DNA methyltransferases (DNMTs) are a large group of enzymes that specifically methylate histone or DNA using S‐adenosyl‐L‐methionine (AdoMet) as a methyl donor. However, there is no widely accepted HTS assay formats and reference inhibitors are available. We have developed and successfully commercialized an extremely low cost miniaturized radioisotope reaction system, the HotSpot SM platform, for kinase screening and profiling to provide services in the drug discovery community. We have effectively adapted this HotSpot SM platform for methyltransferase assays. In this gold standard radioisotope format, tritium‐labeled AdoMet ( 3 H‐AdoMet) was used as a methyl donor and total methylations on substrate were read‐out, which gave high signal/background ratio resulted in high Z’ values. Here, we demonstrate HMTs and DNMTs profiling, kinetic studies, mode of inhibition studies, and HTS can be performed with this platform, which will become a low cost high productive service product to help the drug discovery activities in the emerging epigenetics field. This work was funded in part by NIH SBIR grants R43CA139621, and NIH grants R01HG003818 to H.M.