Table 1.

ITC analyses of binding between AcrIF7 and Cas8f–Cas5f mutants^a

AcrIF7	Cas8f–Cas5f	K D (nM)	N	ΔG (kcal/mol)	ΔH (kcal/mol)	TΔS (kcal/mol)
WT	WT	46 ± 14	0.78 ± 0.00	–10.0 ± 0.2	–17.1 ± 0.2	–7.1 ± 0.3
D13K	WT	2451 ± 528	0.72 ± 0.02	–7.7 ± 0.1	–14.2 ± 0.6	–6.5 ± 0.7
E18K	WT	49 ± 4	0.88 ± 0.00	–10.0 ± 0.1	–19.6 ± 0.1	–9.6 ± 0.1
E22K	WT	50 ± 19	0.75 ± 0.01	–10.0 ± 0.2	–15.6 ± 0.3	–5.6 ± 0.4
D28K/D29K	WT	179 ± 25	0.75 ± 0.00	–9.2 ± 0.1	–12.9 ± 0.1	–3.7 ± 0.2
E33K/E34K	WT	4854 ± 1053	0.90 ± 0.04	–7.3 ± 0.1	–9.5 ± 0.6	–2.3 ± 0.6
E46K/E47K	WT	46 ± 11	0.80 ± 0.00	–10.0 ± 0.2	–17.0 ± 0.2	–7.0 ± 0.2
D57K	WT	333 ± 116	0.77 ± 0.02	–8.9 ± 0.2	–12.7 ± 0.4	–3.8 ± 0.4
WT	K29E(Cas8f)	327 ± 43	0.83 ± 0.01	–8.8 ± 0.1	–14.5 ± 0.2	–5.7 ± 0.2
D13K	K29E(Cas8f)	N.B.b	N.B.	N.B.	N.B.	N.B.
E33K/E34K	K29E(Cas8f)	N.B.	N.B.	N.B.	N.B.	N.B.
WT	K248E(Cas8f)	1873 ± 132	0.78 ± 0.01	–7.8 ± 0.0	–16.0 ± 0.2	–8.2 ± 0.2
D13K	K248E(Cas8f)	N.B.	N.B.	N.B.	N.B.	N.B.
D28K/D29K	K248E(Cas8f)	N.B.	N.B.	N.B.	N.B.	N.B.
E33K/E34K	K248E(Cas8f)	N.B.	N.B.	N.B.	N.B.	N.B.
D57K	K248E(Cas8f)	N.B.	N.B.	N.B.	N.B.	N.B.
AcrIF2c	WT	7.2 ± 2.0	0.84 ± 0.00	–11.0 ± 0.2	–22.2 ± 0.1	–11.1 ± 0.2

^aRaw ITC data are provided in Supplementary Figure S5.

^bNo binding: Integrated heats from the measurement were not sufficient to constrain the least squares fit derived from a one-site binding model for the titration.

^cRef. (24).