Table 3.

Adenosine deamination rate constants measured under single-turnover conditions for ADARld E1008Q and AGS mutants on hGlil RNA^a.

Substrate	Enzyme	k obs (min −1 ) b	k rel c
Gli1	ADAR1d E1008Q	2.0 ± 0.1	1
	ADAR1d E1008Q, G1007R	-	-
	ADAR1d E1008Q, K999N	0.30 ± 0.03	0.1
	ADAR1d E1008Q, R892H	0.02 ± 0.01	0.01
	ADAR1d E1008Q,Y1112F	0.4 ± 0.1	0.4

^aGli1 substrate sequence is shown in Table S1. ADAR1d E1008Q reactions were carried out with 15 mM Tris–HCl pH 7.5, 26 mM KCl, 40 mM potassium glutamate, 1.5 mM EDTA, 0.003% (v/v) NP-40, 4% (v/v) glycerol, 0.5 mM DTT, 1 μg/mL yeast tRNA, and 0.16 U/μL RNAse inhibitor.

^bk_obs was calculated by fitting fitting product formed at different time points to the equation: [P]_t = α[1 −e^−kobst] where [P]_t is percent edited, α is the end point fitted to 95%, and kobs is the observed rate constant.

^ck_rel = k_obs for mutant/k_obs for ADAR1d E1008Q.