A new affinity gel for the purification of α-carbonic anhdrases.


Sahin A., IŞIK S., ARSLAN O., Supuran C. T. , Guler Ö.

Journal of enzyme inhibition and medicinal chemistry, cilt.30, ss.224-8, 2015 (SCI Expanded İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 30 Konu: 2
  • Basım Tarihi: 2015
  • Doi Numarası: 10.3109/14756366.2014.912215
  • Dergi Adı: Journal of enzyme inhibition and medicinal chemistry
  • Sayfa Sayıları: ss.224-8

Özet

The new affinity gel reported in this study was prepared using EUPERGIT C250L as a chromatographic bed material, to which etylenediamine spacer arms were attached to prevent steric hindrance between the matrix and ligand, and to facilitate effective binding of the CA-specific ligand, of the aromatic sulfonamide type for the purification of alpha-carbonic anhydrases (Cas; EC 4.2.1.1). Indeed, the aminoethyl moieties of the affinity gel were derivatized by reaction with 4-isothiocyanatobenzenesulfonamide, with the formation of a thiourea-based gel, having inhibitory effects against CAs. Both bovine erythrocyte carbonic anhydrase BCA and human (h) erythrocyte CA isoforms I, II (hCA I and II) have been purified from hemolysates, by using this affinity gel. The greatest purification fold and column yields for BCA and for cytosolic (hCA I + II) enzymes were of 181-fold (21.07%) and 184-fold (9.49%), respectively. Maximum binding was achieved at 15 degrees C and I = 0.3 ionic strength for alpha-carbonic anhydrases.