6His-Eco29kI methyltransferase methylation site and kinetic mechanism characterization

作者：Nikitin Dmitri^*; Mokrishcheva Marina; Solonin Alexander

来源：Biochimica et Biophysica Acta-Proteins and Proteomics, 2007, 1774(8): 1014-1019.

DOI：10.1016/j.bbapap.2007.05.014

• DNA methyltransferase
• methylation site detection
• kinetic mechanism
• substrate inhibition

A new type 11 6His-Eco29kI DNA methyltransferase was tested for methylation site (CC(Me)GCGG) and catalytic reaction optimal conditions. With high substrate concentrations, an inhibitory effect of DNA, but not AdoMet, on its activity was observed. Isotope partitioning and substrate preincubation assays showed that the enzyme-AdoMet complex is catalytically active. Considering effect of different concentrations of DNA and AdoMet on initial velocity, ping-pong mechanisms were ruled out. According to data obtained, the enzyme appears to work by preferred ordered bi-bi mechanism with AdoMet as leading substrate.

• 出版日期2007-8