Literature summary for 4.2.1.1 extracted from

Bhatt, A.; Mahon, B.P.; Cruzeiro, V.W.; Cornelio, B.; Laronze-Cochard, M.; Ceruso, M.; Sapi, J.; Rance, G.A.; Khlobystov, A.N.; Fontana, A.; Roitberg, A.; Supuran, C.T.; McKenna, R.
Structure-activity relationships of benzenesulfonamide-based inhibitors towards carbonic anhydrase isoform specificity (2017), ChemBioChem, 18, 213-222 .

Cloned(Commentary)

Cloned (Comment)	Organism
isozyme CA II, recombinant expression of wild-type isozyme CA II and chimeric CA IX mimic mutant isozyme in Escherichia coli strain BL21(DE3)	Homo sapiens
isozyme CA IX, recombinant expression of wild-type isozyme in Escherichia coli strain BL21(DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified isozyme CA II (chimeric CA IX mimic) mutant in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, sitting drop vapor diffusion method, mixing of 300-500 nl of 20 mg/mL of CA II and 41 mg/ml of the CA IX mimic solution with crystallization solution containing 1.6 M sodium citrate, 50 mM Tris-HCl, pH 7.8, in a ratio of 3:5 or 1:1, 25°C, 1 week, X-ray diffraction structure determination and analysis at 1.55-1.69 A resolution (PDB IDS 5SZ0, 5SZ1, 5SZ2, and 5SZ3), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model	Homo sapiens
purified isozyme CA IX in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, X-ray diffraction structure determination and analysis at 1.15-1.78 A resolution (PDB IDS 5SZ4, 5SZ5, 5SZ6, and 5SZ7), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model	Homo sapiens

Crystallization (Comment)

Organism

purified isozyme CA II (chimeric CA IX mimic) mutant in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, sitting drop vapor diffusion method, mixing of 300-500 nl of 20 mg/mL of CA II and 41 mg/ml of the CA IX mimic solution with crystallization solution containing 1.6 M sodium citrate, 50 mM Tris-HCl, pH 7.8, in a ratio of 3:5 or 1:1, 25°C, 1 week, X-ray diffraction structure determination and analysis at 1.55-1.69 A resolution (PDB IDS 5SZ0, 5SZ1, 5SZ2, and 5SZ3), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model

Homo sapiens

purified isozyme CA IX in complex with inhibitors 4-phenylbenzene-1-sulfonamide, 4-(2-methylphenyl)benzene-1-sulfonamide, 4-(3-formylphenyl)benzene-1-sulfonamide, and 4-(3-quinolinyl)benzene-1-sulfonamide, X-ray diffraction structure determination and analysis at 1.15-1.78 A resolution (PDB IDS 5SZ4, 5SZ5, 5SZ6, and 5SZ7), molecular replacement using the enzyme structure PBD ID 3KS3 without zinc and solvent as search model

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
A65S/N67Q/E69T/I91L/F131V/K170E/L204A	site-directed mutagenesis, active site mutant	Homo sapiens
additional information	construction of a chimeric enzyme, the mimic is a chimeric protein of CA II containing seven active-site-residue mutations to resemble CA IX, allowing for ease of protein expression and crystallization. The active-site mutations, A65S, N67Q, E69T, I91L, F131V, K170E, L204A, result in approximately 90% sequence homology between the mimic and wild-type CA IX active sites, such that drugs bound to the CA IXb mimic are representative of the interactions that occur with the wild type enzyme	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
4-(2-methylphenyl)benzene-1-sulfonamide	-	Homo sapiens
4-(3-formylphenyl)benzene-1-sulfonamide	-	Homo sapiens
4-(3-quinolinyl)benzene-1-sulfonamide	-	Homo sapiens
4-phenylbenzene-1-sulfonamide	-	Homo sapiens
acetazolamide	-	Homo sapiens
additional information	structure-activity relationships of benzenesulfonamide-based inhibitors towards carbonic anhydrase isoform specificity, inhibition of different human isozymes, in silico modeling of inhibitors in CA I, oveview; structure-activity relationships of benzenesulfonamide-based inhibitors towards carbonic anhydrase isoform specificity, inhibition of different human isozymes, overview; structure-activity relationships of benzenesulfonamide-based inhibitors towards carbonic anhydrase isoform specificity, inhibition of different human isozymes, overview; structure-activity relationships of benzenesulfonamide-based inhibitors towards carbonic anhydrase isoform specificity, inhibition of different human isozymes, overview	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Homo sapiens	5829	-
extracellular	extracellular membrane-bound	Homo sapiens	-	-
membrane	extracellular membrane-bound	Homo sapiens	16020	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	required, metalloenzyme	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
H2CO3	Homo sapiens	-	CO2 + H2O	-	r

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O43570	-	-
Homo sapiens	P00915	-	-
Homo sapiens	P00918	-	-
Homo sapiens	Q16790	-	-

Purification (Commentary)

Purification (Comment)	Organism
isozyme CA II, recombinant wild-type isozyme CA II and chimeric CA IX mimic mutant isozyme from Escherichia coli strain BL21(DE3) by p-aminomethylbenzenesulfonamide affinity chromatography	Homo sapiens
isozyme CA IX, recombinant wild-type isozyme from Escherichia coli strain BL21(DE3) by p-aminomethylbenzenesulfonamide affinity chromatography	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
H2CO3	-	Homo sapiens	CO2 + H2O	-	r

Synonyms

Synonyms	Comment	Organism
CA I	-	Homo sapiens
CA II	-	Homo sapiens
CA IX	-	Homo sapiens
CA XII	-	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.0000002	-	4-phenylbenzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000002	-	4-(3-quinolinyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000006	-	4-(3-formylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000008	-	4-phenylbenzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000012	-	4-(2-methylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000002	-	4-(3-formylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000025	-	4-(3-quinolinyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0000057	-	acetazolamide	pH and temperature not specified in the publication	Homo sapiens
0.000012	-	acetazolamide	pH and temperature not specified in the publication	Homo sapiens
0.00002	-	4-(2-methylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000025	-	acetazolamide	pH and temperature not specified in the publication	Homo sapiens
0.000035	-	4-(3-quinolinyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000042	-	4-phenylbenzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000047	-	4-(2-methylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000093	-	4-phenylbenzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.000099	-	4-(3-formylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.00025	-	acetazolamide	pH and temperature not specified in the publication	Homo sapiens
0.00076	-	4-(3-quinolinyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0012	-	4-(3-formylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens
0.0035	-	4-(2-methylphenyl)benzene-1-sulfonamide	pH and temperature not specified in the publication	Homo sapiens