A nonradioactive DNA methyltransferase assay adaptable to high-throughput screening

Youn Hi Woo, P. T. Ravi Rajagopalan, Stephen J. Benkovic

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

We have developed a nonradioactive assay method for DNA methyltransferases based on the ability to protect substrate DNA from restriction. DNA immobilized to a microplate well was treated sequentially with methyltransferase and an appropriate endonuclease. The amount of methylated DNA product is reflected by a proportional decrease in endonuclease cleavage, which is in turn reflected by increased retention of the end-labeled affinity probe. A single universal substrate was designed to assay multiple methyltransferases including those that do not have a cognate endonuclease. The methodology developed is suited to screen a large number of compounds for inhibitors of various methyltransferases.

Original languageEnglish (US)
Pages (from-to)336-340
Number of pages5
JournalAnalytical Biochemistry
Volume340
Issue number2
DOIs
StatePublished - May 15 2005

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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