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

BMC Biochemistry

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

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.	0.7^b	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.	1.20^b	<0.100	12	0.9

^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
The given values are representative of two or more experiments
^bData from ref. [24]

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