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

Demirdag R., Comakli V., Senturk M., Ekinci D., KÜFREVİOĞLU Ö. İ., SUPURAN C. T.

BIOORGANIC & MEDICINAL CHEMISTRY, cilt.21, sa.6, ss.1522-1525, 2013 (SCI-Expanded) identifier identifier identifier

  • 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
  • Atatürk 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.