title: Purification and characterization of carbonic anhydrase from Ağrı Balık Lake trout gill (Salmo trutta labrax) and effects of sulfonamides on enzyme activity.
abstract: Carbonic anhydrase (CA) was purified from Ağrı Balık Lake trout gill (fCA) by affinity chromatography on a sepharose 4B-tyrosine-sulfanilamide column. The fCA enzyme was purified with about a 303.9 purification factor, a specific activity 4130.4 EU (mg-protein)(-1), and a yield of 79.3 by using sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography. The molecular weight determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was found to be about 29.9 kDa. The kinetic parameters, K(M) and V(max) were determined for the 4-nitrophenyl acetate hydrolysis reaction. Some sulfonamides were tested as inhibitors against the purified CA enzymes. The Ki constants for mafenide (1), p-toluenesulfonamide (2), 2-bromo-benzene sulfonamide (3), 4-chlorobenzene sulfonamide (4), 4-amino-6-chloro-1-3 benzenedisulfonamide (5), sulfamethazine (6), sulfaguanidine (7), sulfadiazine (8), and acetozazolamide (9) were in the range of 7.5-108.75 μM.
authors.affiliation: School of Health, Agri Ibrahim Cecen University, Agri, Turkey. r.demirdag@hotmail.com.
authors.forename: Ramazan
authors.initials: R
authors.lastname: Demirdag
authors.forename: Veysal
authors.initials: V
authors.lastname: Comakli
authors.forename: Muslum
authors.initials: M
authors.lastname: Kuzu
authors.forename: Emrah
authors.initials: E
authors.lastname: Yerlikaya
authors.forename: Murat
authors.initials: M
authors.lastname: Şentürk
mesh_terms: D000818:Animals; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D005880:Gills / Q000201:enzymology*; D008970:Molecular Weight; D013424:Sulfanilamides / Q000494:pharmacology*; D014337:Trout / Q000378:metabolism*
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D013424:Sulfanilamides; D002256:Carbonic Anhydrases
keywords: Ağrı Balık Lake Trout; Carbonic Anhydrase; Characterization; Inhibition; Purification; Sulfonamides