Table 2.

Calculated Free Energy Profile for the Catalytic Reaction of Cas9 Different Systems^a

ref				LYS848 far			LYS848 near/ionized			LYS848 near/un-ionized
ΔGpt	Δg‡,NA	Δg‡	$\Delta g_{\text{obs }}^{‡}$	ΔGpt	Δg‡,NA	Δg‡	ΔGpt	Δg‡,NA	Δg‡	ΔGpt	Δg‡,NA	Δg‡	$\Delta g_{\text{obs }}^{{}^{‡}\text{WT}}$
10.93	33	43.93	43.93	27.49	20.73	48.22	3.64	17.37	21.01	4.38	21.97	26.35	17.80

^aCalculated free energies (kcal/mol) of proton transfer (PT) step, nucleophilic attack (NA) step, and total barrier in the reference solution and the three cases for Cas9. The observed barrier was converted from the k_cat of ref 26.