Skip to main content

Table 2 Methylation activity, DNA binding affinity, and 2-aminopurine base flipping by Dnmt3a-CD mutantsa

From: Conserved motif VIII of murine DNA methyltransferase Dnmt3a is essential for methylation activity

Exchange Corresponding residue in full length Dnmt3a K d1, nM K d2, nM K d1 K d2, nM2 Relative binding affinity v 0, nM/min v 0 rel, % Relative intensity of 2-aminopurine fluorescence, at 370 nm, %
wt   73 ± 17 14 ± 5 1022 1.00 0.83 ± 0.09 100 100
N167A N753 n.d. n.d. n.d. 0.7b 0.14 ± 0.02 17 29.4
R200A R786 79 ± 17 14 ± 6 1106 0.92 <0.020 2 3.9
R200E R786 159 ± 37 33 ± 12 5247 0.19 <0.022 3 5.3
R200K R786 55 ± 14 9 ± 3 495 2.06 <0.035 4 15.4
R202A R788 n.d. n.d. n.d. 1.20b <0.100 12 0.9
  1. aThe amino acid numbers in Dnmt3a catalytic domain are given relative to the murine full-length Dnmt3a (NCBI reference sequence NP_001258682.1). n.d. designates not determined
  2. The given values are representative of two or more experiments
  3. bData from ref. [24]