title: Catalysis by mutants of human carbonic anhydrase II: effects of replacing hydrophobic residues 198 and 204.
abstract: Previous studies shows that the replacement of Phe-198 in carbonic anhydrase III to the corresponding Leu residue found in carbonic anhydrase II caused the appearance of isozyme II-like activity (LoGrasso et al. (1991) Biochemistry 30, 8463-8470). Carbonic anhydrase II is more efficient in the catalysis of CO2 hydration by 500-fold and has an apparent pKa for this catalysis about two pKa units above that of carbonic anhydrase III. Moreover, isozyme II catalyzes the hydrolysis of 4-nitrophenyl acetate, whereas isozyme III shows no appreciable catalysis. The purpose of this work was to test the hypothesis that making the converse replacement Leu-198-->Phe as well as Leu-204-->Glu and the double replacement in carbonic anhydrase II would give the resulting mutants of isozyme II properties of isozyme III. The catalytic activities of these mutants in CO2 hydration and 4-nitrophenyl acetate hydrolysis were smaller by at most 5-fold and the pKa values for these catalyses were identical compared with wild-type isozyme II. The different effects of converse mutants of HCA II and III indicate complexity in structure not evident from their similar backbone conformations.
authors.affiliation: Department of Pharmacology, University of Florida, Gainesville 32610-0267.
authors.forename: S
authors.initials: S
authors.lastname: Taoka
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: R W
authors.initials: RW
authors.lastname: Tarnuzzer
authors.forename: G
authors.initials: G
authors.lastname: Van Heeke
authors.forename: C
authors.initials: C
authors.lastname: Tu
authors.forename: D N
authors.initials: DN
authors.lastname: Silverman
mesh_terms: D001483:Base Sequence; D001639:Bicarbonates / Q000378:metabolism; D001692:Biological Transport; D002245:Carbon Dioxide / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000502:physiology; D002384:Catalysis; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011522:Protons; D013329:Structure-Activity Relationship
chemical_list: D001639:Bicarbonates; D011522:Protons; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases