Table 1. Impact on CacKam activity and ligand interactions of residue substitutions around the SAM-binding pocket.

Amino acid substitution	Kanamycin MIC (μg/ml)	Ligand bindinga,b
		SAM Kd (μM)	SAH Kd (μM)
-	>1024	44	2.7
D36A	8	NB	NB
D61A	8	NB	NB
R66A	512	21	2.9
E94A	16	29	2.2
K115A	256	48	5.0
S201A	16	110	1.6
S202A	8	24	2.9
R43A/R73A	16	35	2.4
W113A	16	31	1.9
W113F	16	103	11
W203A	16	68	5.1
W203F	16	24	1.9

^aData for wild-type CacKam are from reference (19).

^bNB, denotes no binding detected.