Purification and characterization of carbonic anhydrase from sheep kidney and effects of sulfonamides on enzyme activity.

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Demirdag R, Comakli V, Senturk M, Ekinci D, Irfan Kufrevioglu O, Supuran CT

Purification and characterization of carbonic anhydrase from sheep kidney and effects of sulfonamides on enzyme activity.

Bioorg Med Chem. 2013 Mar 15;21(6):1522-5. doi: 10.1016/j.bmc.2012.08.018. Epub 2012 Aug 24.

PubMed ID

22974493 [ View in PubMed

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Abstract

Carbonic anhydrase (CA, EC: 4.2.1.1) was purified from sheep kidney by affinity chromatography on a Sepharose 4B-tyrosine-sulfanilamide column. By means of two consecutive procedures, the enzyme (sCA) was purified 227.61-fold with a yield of 60.75%, and a specific activity of 838.89U/mg proteins. The optimum temperature, ionic strength and pH were determined to be 35 degrees C, 20mM and 8.5, respectively. The molecular weight determined by SDS-PAGE was found to be 29kDa. The kinetic parameters, KM and Vmax values were determined for the 4-nitrophenyl acetate (p-NpA) hydrolysis reaction. Some sulfonamides were tested as inhibitors against the purified CAs enzyme. The Ki constants for benzenesulfonamide (1), sulfanilamide (2), mafenide (3), 4-(2-aminoethyl) benzenesulfonamide (4), 4-methyl-benzenesulfonamide (5), 2-bromo-benzenesulfonamide (6), naphthalene-2-sulfonamide (7), 4-amino-6-chlorobenzene-1,3-disulfonamide (8) and saccharin (9) were in the range 1.348-69.31muM.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Mafenide	Carbonic anhydrase 6	Protein	Humans	Unknown	Antagonist	Details

Binding Properties

Drug	Target	Property	Measurement	pH	Temperature (°C)
4-(2-AMINOETHYL)BENZENESULFONAMIDE	Carbonic anhydrase 2	Ki (nM)	160	N/A	N/A	Details