Table 6. Comparison of the Final Results from Simulation and Experiment for Both Sites of Point Mutation and Different P1′ Amino Acids^a.

	mutation at 105				mutation at 171
	P1’ = Ile		P1’ = Pro		P1’ = Ile		P1’ = Pro
	sim	exp	sim	exp	Sim	exp	sim	exp
Ala	–19.9	√	–1.2	√	–14.3	√	11.5	√
Arg	–2.5	√	–14.7		–4.1	√	4.9	√
Asn	–14.1	√	–16.7		–0.8		–1.8	√
Asp	1.1		6.9		–10.3		–3.9	√
Cys	–13.8	√	–12.2		5.4	√	0.7	√
Gly	–1.8	√	0.1		0		0
Gln	–5.5	√	11.8		–8.4		–2.1	√
Glu	0	√	0		–9.9	√	–8.2	√
HisA	–14.7	√	–1.1		0.9		–4.5
HisB	–16.5	√	–3.6		3.4		10.9
Ile	–9.7	√	6.1		0.9		–10.0
Leu	–10.7	√	7.2		0.3		–7.8
Lys	–11.4		16.1		–4.5	√	5.3
Met	–6.2	√	–12.3	√	0.8		8.3
Phe	1.5		5.3		3.0		6.7
Ser	–22.2	√	12.9		–3.7	√	6.3
Thr	–11.9	√	11.5		–6.6		0.2
Trp	–79.5	√	–18.2		–1.2	√	3.8
Tyr	–9.4	√	9.0		16.8		–0.3
Val	–9.7	√	0.0	√	–8.7	√	–7.1	√

^aRelative substrate binding free energies (“sim” column) are reported for the mutations Glu105Xxx and Gly171Xxx (relative to the unmated protein). In the columns with the experimental results, check marks/horizontal bars mark mutations that were found/not found in the screening. All values are reported in kJ/mol. HisA: N^δ1–H tautomer; HisB: N^ϵ2–H tautomer.