Analysis of the mechanism of the Serratia nuclease using site-directed mutagenesis.

  title={Analysis of the mechanism of the Serratia nuclease using site-directed mutagenesis.},
  author={Peter Friedhoff and B Kolmes and Oleg A. Gimadutdinow and Wolfgang Wende and Kurt L Krause and Alfred Pingoud},
  journal={Nucleic acids research},
  volume={24 14},
Based on crystal structure analysis of the Serratia nuclease and a sequence alignment of six related nucleases, conserved amino acid residues that are located in proximity to the previously identified catalytic site residue His89 were selected for a mutagenesis study. Five out of 12 amino acid residues analyzed turned out to be of particular importance for the catalytic activity of the enzyme: Arg57, Arg87, His89, Asn119 and Glu127. Their replacement by alanine, for example, resulted in mutant… CONTINUE READING
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