Sulphonamide inhibition profile of Staphylococcus aureus β-carbonic anhydrase

Figures & data

Figure 1. SDS-PAGE of the purified SauBCA with and without the His-tag. The polypeptides marked with arrows were subjected to MS/MS analysis and identified as SauBCA.

Table 1. Kinetic data of SauBCA and the inhibition results for the standard sulphonamide inhibitor acetazolamide (AAZ). All human isozymes^22,48 and two other β-CAs^26,27,49 with corresponding properties are shown for comparison. The k_cat and k_cat/K_M values are rounded to one decimal place.

Enzyme	kcat	kcat/KM	Ki (AAZ)
	(s-1)	(s-1M-1)	(nM)
SauBCA	1.5 × 10^5	2.6 × 10^7	628
hCA I	2.0 × 10^5	5.0 × 10^7	250
hCA II	1.4 × 10^6	1.5 × 10^8	12
hCA III	1.3 × 10^4	2.5 × 10^5	240000
hCA IV	1.1 × 10^6	5.1 × 10^7	74
hCA VA	2.9 × 10^5	2.9 × 10^7	63
hCA VB	9.5 × 10^5	9.8 × 10^7	54
hCA VI	3.4 × 10^5	4.9 × 10^7	11
hCA VII	9.5 × 10^5	8.3 × 10^7	2.5
hCA IX	1.1 × 10^6	1.5 × 10^8	16
hCA XII	4.2 × 10^5	3.5 × 10^7	5.7
hCA XIII	1.5 × 10^5	1.1 × 10^7	16
hCA XIV	3.1 × 10^5	3.9 × 10^7	41
Burkholderia pseudomallei β-CA	1.6 × 10^5	3.4 × 10^7	745
Flaveria bidentis β-CA	1.2 × 10^5	7.5 × 10^6	27

Figure 2. The molecular structures of the sulphonamide analogues used in this study (1–24) as well as selected clinically used agents.

Table 2. Inhibition data for SauBCA and hCA II with sulphonamide analogues 1–24 and selected clinically used agents.

Ki* (nM)
Inhibitor	SauBCA	hCA II^a
1	355	300
2	409	240
3	95	8
4	83	320
5	193	170
6	253	160
7	93	60
8	95	110
9	75	40
10	202	54
11	81	63
12	79	75
13	417	60
14	553	19
15	619	80
16	603	94
17	232	125
18	555	46
19	909	33
20	92	2
21	85	11
22	83	46
23	92	33
24	96	30
AAZ	628	12
MZA	863	14
EZA	698	8
DZA	909	9
BRZ	815	3
BZA	501	9
TPM	466	10
ZNS	4551	35
SLP	807	40
IND	588	15
VLX	509	43
CLX	871	21
SLT	824	9
SAC	667	5959
HCT	593	290
FAM	71	58
EPA	538	917

*Mean from three different assays measured by the stopped-flow technique. Errors were in the range of ±5–10% of the reported values (data not shown). ^aHuman recombinant isozyme, from Ref.²².

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