Whitney Shatz
Research Abstract
Enzymatic DNA methylation is involved in the regulation of gene expression. Mammalian DNA methyltransferases have been identified as targets for anti-cancer drugs. To further understand the mechanism of DNA methyltransferases, the correlation between active site and non-active site motions of M.HhaI, a bacterial homolog, is being studied. A Trosy solution state NMR spectrum has been obtained with 15N labeled protein, but due to its 37 kDa size the signal to noise ratio is low. Higher concentrations are needed to increase the signal strength and sharpen the broad peaks. At such high concentrations, the wild type protein is weakly soluble and dimerizes. Different combinations of DNA and cofactor, varying buffer composition and the engineering of new constructs are being explored to overcome these issues in order to obtain a triply labeled Trosy spectrum necessary for making residue assignments.
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