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. Author manuscript; available in PMC: 2020 Oct 21.
Published in final edited form as: J Med Chem. 2019 May 29;62(11):5616–5627. doi: 10.1021/acs.jmedchem.9b00561

Table 2.

Displacement assays, thermal shift data for aryl-sulfonyl fluorides and aryl-fluoro sulfates tested against several XIAP BIR3 mutants.

graphic file with name nihms-1636519-t0007.jpg
Agent BIR3wt Lys311Ala Lys311Tyr Lys311His Lys311Thr Lys311Ser
ID X IC50a
[nM]		 ΔTm
[°C]b 		IC50
[nM]a 		ΔTm
[°C]b 		IC50
[nM]a 		ΔTm
[°C]b 		IC50
[nM]a 		ΔTm
[°C]b 		IC50
[nM]a 		ΔTm
[°C]b 		IC50
[nM]a 		ΔTm
[°C]b
2 graphic file with name nihms-1636519-t0008.jpg 11 ± 1
13 ± 1		 31
37		 86 ± 2
59 ± 6		 1.5
8.5		 12 ± 2
10 ± 0.8		 37.5
34.5		 26 ± 0.1
21 ± 1		 35.5
28.5		 201 ± 1
72 ± 2		 4.0
6.5		 160 ± 8
80 ± 9		 5
11
3 graphic file with name nihms-1636519-t0009.jpg 17 ± 0.1
8 ± 0.1		 35.5
35		 61 ± 4
27 ± 0.1		 5
7.5d 		6.4 ± 0.1
3.5 ±0.1		 36
33		 62 ± 14
16 ± 1		 31
28.5		 161 ± 9
45 ± 2		 5
6d 		145 ± 7
39 ± 1		 5
7.5d
4 graphic file with name nihms-1636519-t0010.jpg 71 ± 7
24 ± 1		 8.5
33.0		 65 ± 1
55 ± 0.2		 11
13.5		 37 ± 5
21 ± 2		 8.5
35.5		 113± 22
63 ± 4		 11
12.5		 140±20
120±10		 9.5
11.5		 177 ± 4
126 ± 2		 10.5
14.5
5 graphic file with name nihms-1636519-t0011.jpg 190± 20
105± 10		 11.5
16		 86 ± 9
95 ± 4		 10.5
11.5		 55± 16
71 ± 8		 10.5
11.5		 137 ± 4
110 ± 3		 8.5
8.5		 231±20
220±10		 8.5
9.5		 241 ± 1
230± 10		 6.5
13.5
6 graphic file with name nihms-1636519-t0012.jpg 140 ± 1
100 ± 5		 11.5
15.0		 89 ± 0.2
108 ± 6		 10.5
11.5		 63 ± 10
85 ± 4		 10.5
11.5		 140± 24
130± 10		 11.5
12		 240±12
220±20		 7.5
9.5		 250±1
276±14		 8
13.5
7 graphic file with name nihms-1636519-t0013.jpg 590± 65
100 ± 7		 4.5
31c 		218 ±8
270± 25		 8
8.5		 113 ± 2
48 ± 6		 5.5
32		 270±70
260±20		 36.5e
38e 		620±40
630±20		 5.5
6.5		 713± 3
646± 32		 8.5
10.5
8 graphic file with name nihms-1636519-t0014.jpg 2000± 20
90 ± 1		 11.5
28.5c 		70 ± 2
70 ± 7		 11
12.5		 80 ± 2
86 ± 13		 10.5
29.5c 		126± 14
98 ± 1		 11
29.5c 		180± 40
165± 3		 8.5
10		 250± 1
237± 13		 10.5
10.5
a

IC50 values represent dose-response curves obtained after incubating protein and ligand for 2 h or 8 h at room temperature.

b

Thermal shift (ΔTm) data were obtained incubating protein and ligand for 2 h at room temperature or 6 h at 37°C. The data reported represent the major peak observed (see supplementary information for individual curves).

c

In this experiment, the peak corresponding to the non-covalent ΔTm is still present.

d.

An additional smaller peak that corresponds to a ΔTm ~30 °C is observed perhaps due to a covalent interaction with Lys322.

e.

In this experiment the shift reported corresponds to a minor peak.