TY - JOUR
T1 - A nonradioactive DNA methyltransferase assay adaptable to high-throughput screening
AU - Woo, Youn Hi
AU - Ravi Rajagopalan, P. T.
AU - Benkovic, Stephen J.
PY - 2005/5/15
Y1 - 2005/5/15
N2 - 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.
AB - 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.
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U2 - 10.1016/j.ab.2005.01.059
DO - 10.1016/j.ab.2005.01.059
M3 - Article
C2 - 15840507
AN - SCOPUS:17444389293
SN - 0003-2697
VL - 340
SP - 336
EP - 340
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -