Cytosolic and non-cytosolic carbonic anhydrase enzymes from bovine leukocytes.
In this study, carbonic anhydrase (CA) enzyme has been purified and separately characterized according to bound form in 4 steps as outer peripheral, cytosolic, inner peripheral, and integral from bovine leukocyte. Affinity chromatography has also been used for purification of the enzyme in four steps. CA has been found for each step. Measurment of enzyme activity has been done by CO2 hydratase activity and esterase activity methods. Optimum pH and optimum temperature have been defined for each step of purified enzyme. The behaviors of CA with specific inhibitors, such as KSCN and NaN3 have been investigated. In each step, molecular weight and purity have been determined by gel filtration and SDS-PAGE electrophoresis. In addition, enzyme K(M) and Vmax values have been determined with the method of Lineweaver-Burk.
Demir
Y
Y
Ataturk Universitesi, Fen-Edebiyat Fakultesi, Kimya Bolumu, Erzurum, Turkey.
Demir
N
N
Alayli
A
A
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D002256:Carbonic Anhydrases / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods; D003600:Cytosol / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007962:Leukocytes / Q000201:enzymology*; D009584:Nitrogen / Q000494:pharmacology; D013696:Temperature; D013861:Thiocyanates / Q000494:pharmacology
D000963:Antimetabolites; D004791:Enzyme Inhibitors; D013861:Thiocyanates; C003320:sodium nitride; D002256:Carbonic Anhydrases; D009584:Nitrogen; C009941:potassium thiocyanate