PMID: 2114290Jun 20, 1990Paper

Fine tuning of the catalytic properties of carbonic anhydrase. Studies of a Thr200----His variant of human isoenzyme II

European Journal of Biochemistry
G BehravanS Lindskog

Abstract

The active sites of carbonic anhydrases I contain a unique histidine residue at sequence position 200. To test the hypothesis that His200 is essential for the isoenzyme-specific catalytic and inhibitor-binding properties of carbonic anhydrases I, a variant of human carbonic anhydrase II, having His200 for Thr200, was prepared by oligonucleotide-directed mutagenesis. The variant has a circular dichroic spectrum that is indistinguishable from that of the parent enzyme. The kinetics of CO2 hydration and HCO3- dehydration has been investigated. The results show that the amino acid substitution has led to changes of catalytic parameters as well as Ki values for anion inhibition in the expected directions towards the values for isoenzyme I. However, the maximal 4-nitrophenyl acetate hydrolase activity of the variant is higher than for any naturally occurring carbonic anhydrase studied so far. A detailed analysis of the kinetic observations suggests that the modification has resulted in a change of the step that limits the maximal rate of CO2 hydration at saturating buffer concentrations. This rate-limiting step is an intramolecular proton transfer in unmodified isoenzyme II and, presumably, HCO3- dissociation in the variant and in hu...Continue Reading

References

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May 1, 1988·Acta Chemica Scandinavica. Series B: Organic Chemistry and Biochemistry·C ForsmanB H Jonsson
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Citations

Jan 1, 1997·Pharmacology & Therapeutics·S Lindskog
May 9, 2001·Biochimica Et Biophysica Acta·L C ChiricaS Lindskog
Mar 4, 2000·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·A Scozzafava, C T Supuran
Nov 13, 2003·Bioorganic & Medicinal Chemistry Letters·Marco FranchiClaudiu T Supuran
Apr 20, 2001·Bioorganic & Medicinal Chemistry·C T SupuranA Scozzafava
Apr 26, 2003·Bioorganic & Medicinal Chemistry·Andrea Scozzafava, Claudiu T Supuran
Oct 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Vincenzo AlterioGiuseppina De Simone
Aug 18, 2005·Proteins·Deepa BhattDavid N Silverman
Feb 3, 2005·Bioorganic & Medicinal Chemistry Letters·Alessio InnocentiClaudiu T Supuran
Dec 1, 1993·European Journal of Biochemistry·I M Johansson, C Forsman
Jan 15, 1994·European Journal of Biochemistry·A LiljasY Xue
May 1, 1995·European Journal of Biochemistry·C EngstrandS Lindskog
Jan 18, 2006·Chemistry : a European Journal·Krzysztof Okrasa, Romas J Kazlauskas
Oct 9, 2014·Molecular BioSystems·Manish Kumar TiwariJung-Kul Lee
Oct 12, 2018·International Journal of Medicinal Chemistry·Carrie L LomelinoRobert McKenna
Dec 25, 2003·Journal of Enzyme Inhibition and Medicinal Chemistry·Daniela VulloClaudiu T Supuran
Jan 11, 2002·Journal of Medicinal Chemistry·Bernard MasereelClaudiu T Supuran

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