Characterization and anions inhibition studies of an α-carbonic anhydrase from the teleost fish Dicentrarchus labrax.

Ekinci D., CEYHUN S. B., SENTURK M., ERDEM D., KÜFREVİOĞLU Ö. İ., SUPURAN C. T.

Bioorganic & medicinal chemistry, cilt.19, sa.2, ss.744-8, 2011 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 19 Sayı: 2
  • Basım Tarihi: 2011
  • Doi Numarası: 10.1016/j.bmc.2010.12.033
  • Dergi Adı: Bioorganic & medicinal chemistry
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.744-8
  • Anahtar Kelimeler: Carbonic anhydrase, Alpha-class, Teleost fish, Esterase, Anion inhibitor, Sulfamic acid, Thiocyanate, IN-VITRO, ISOZYME-I, OPERCULAR EPITHELIUM, ESTERASE-ACTIVITIES, FRESH, TRANSMEMBRANE, WATER, XIII
  • Atatürk Üniversitesi Adresli: Evet

Özet

Carbonic anhydrase (CA; EC 4.2.1.1) was purified from the gill of the teleost fish Dicentrarchus labrax (European seabass). The purification procedure consisted of a single step affinity chromatography on Sepharose 4B-tyrosine-sulfanilamide. The enzyme was purified 84.9-fold with a yield of 58%, and a specific activity of 838.9 U/mg proteins. It has an optimum pH at 8.0; an optimum temperature at 10 degrees C. The kinetic parameters of this enzyme were determined for its esterase activity, with 4-nitrophenyl acetate (NPA) as substrate. The following anions, H2NSO3-, I , SCN , NO3- , NO2- , N-3(-) , Br , Cl , SO42- , and F showed inhibitory effects on the enzyme. Sulfamic acid, iodide, and thiocyanate exhibited the strongest inhibitory action, in the micromolar range (K(i)s of 87-187 mu M). NO3- , NO2- and N-3(-) were moderate inhibitors, whereas other anions showed only weak actions. All tested anions inhibited the enzyme in a competitive manner. Our findings indicate that these anions inhibit the fish enzyme in a similar manner to other a-CAs from mammals investigated earlier, but the susceptibility to various anions differs significantly between the fish and mammalian CAs. (C) 2010 Elsevier Ltd. All rights reserved.