Purification and characterization of carbonic anhydrase from sheep kidney and effects of sulfonamides on enzyme activity
BIOORGANIC & MEDICINAL CHEMISTRY, cilt.21, sa.6, ss.1522-1525, 2013 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 21 Sayı: 6
- Basım Tarihi: 2013
- Doi Numarası: 10.1016/j.bmc.2012.08.018
- Dergi Adı: BIOORGANIC & MEDICINAL CHEMISTRY
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Sayfa Sayıları: ss.1522-1525
- Anahtar Kelimeler: Carbonic anhydrase, Sheep, Kidney enzyme, Inhibition, Sulfonamides, IN-VITRO INHIBITION, SEA-BASS LIVER, DERIVATIVES, ISOZYMES, PESTICIDES, ESTERASE, LABRAX, SITE, VI
- Ondokuz Mayıs Üniversitesi Adresli: Evet
Özet
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 838.89 U/mg proteins. The optimum temperature, ionic strength and pH were determined to be 35 degrees C, 20 mM and 8.5, respectively. The molecular weight determined by SDS-PAGE was found to be 29 kDa. The kinetic parameters, K-M and V-max values were determined for the 4-nitrophenyl acetate (p-NpA) hydrolysis reaction. Some sulfonamides were tested as inhibitors against the purified CAs enzyme. The K-i 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 1.348-69.31 mu M. (C) 2012 Elsevier Ltd. All rights reserved.