Direct identification of the active-site nucleophile in a DNA (cytosine-5)-methyltransferase.

  title={Direct identification of the active-site nucleophile in a DNA (cytosine-5)-methyltransferase.},
  author={Liwei Chen and Andrew M. MacMillan and W Chang and Khosro Ezaz-Nikpay and William S. Lane and Gregory L. Verdine},
  volume={30 46},
The overproduction, purification, and determination of the active-site catalytic nucleophile of the DNA (cytosine-5)-methyltransferase (DCMtase) enzyme M.HaeIII are reported. Incubation of purified M.HaeIII with an oligodeoxynucleotide specifically modified with the mechanism-based inhibitor 5-fluoro-2'-deoxycytidine [Osterman, D. G., et al. (1988) Biochemistry 27, 5204-5210], in the presence of the cofactor S-adenosyl-L-methionine (AdoMet), resulted in the formation of a covalent DNA-M.HaeIII… CONTINUE READING
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