PMID: 11341916May 9, 2001Paper

Cloning, expression and some properties of alpha-carbonic anhydrase from Helicobacter pylori

Biochimica Et Biophysica Acta
L C ChiricăS Lindskog

Abstract

The alpha-carbonic anhydrase gene from Helicobacter pylori strain 26695 has been cloned and sequenced. The full-length protein appears to be toxic to Escherichia coli, so we prepared a modified form of the gene lacking a part that presumably encodes a cleavable signal peptide. This truncated gene could be expressed in E. coli yielding an active enzyme comprising 229 amino acid residues. The amino acid sequence shows 36% identity with that of the enzyme from Neisseria gonorrhoeae and 28% with that of human carbonic anhydrase II. The H. pylori enzyme was purified by sulfonamide affinity chromatography and its circular dichroism spectrum and denaturation profile in guanidine hydrochloride have been measured. Kinetic parameters for CO2 hydration catalyzed by the H. pylori enzyme at pH 8.9 and 25 degrees C are kcat=2.4x10(5) s(-1), KM=17 mM and kcat/KM=1.4x10(7) M(-1) x s(-1). The pH dependence of kcat/KM fits with a simple titration curve with pK(a)=7.5. Thiocyanate yields an uncompetitive inhibition pattern at pH 9 indicating that the maximal rate of CO2 hydration is limited by proton transfer between a zinc-bound water molecule and the reaction medium in analogy to other forms of the enzyme. The 4-nitrophenyl acetate hydrolase ac...Continue Reading

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Citations

Mar 11, 2003·Nucleic Acids Research·Ivo G BonecaIvan Moszer
Feb 17, 2009·Journal of Bacteriology·Chrysanthi KalloniatiEmmanouil Flemetakis
Aug 24, 2002·Journal of Bacteriology·Bernhard KusianBotho Bowien
Nov 18, 2008·Journal of Gastroenterology·Saori MorishitaClaudiu T Supuran
Nov 26, 2002·Biochimica Et Biophysica Acta·Laura C ChiricaSven Lindskog
Mar 8, 2011·FEBS Letters·Jose Antonio Cuesta-SeijoLeila Lo Leggio
Oct 27, 2004·Journal of Enzyme Inhibition and Medicinal Chemistry·Silvia PastorekovaClaudiu T Supuran
Jul 19, 2012·Journal of Enzyme Inhibition and Medicinal Chemistry·Clemente CapassoMosè Rossi
Jan 25, 2011·Biochimica Et Biophysica Acta·Daniela TsikouEmmanouil Flemetakis
Mar 12, 2004·Bioorganic & Medicinal Chemistry Letters·Silvia PastorekovaClaudiu T Supuran
Jan 28, 2015·Antimicrobial Agents and Chemotherapy·Dhiman Sankar PalRupak Datta
Dec 5, 2019·Environmental Science and Pollution Research International·Anand Giri, Deepak Pant
Nov 7, 2001·The Journal of Biological Chemistry·B C TrippJ G Ferry
Feb 13, 2008·Biochemistry·Albert A BarreseBrian C Tripp
Mar 17, 2019·International Journal of Biological Macromolecules·Neha MaheshwariShaili Srivastava
Mar 20, 2003·Bioorganic & Medicinal Chemistry Letters·Daniela VulloClaudiu T Supuran
Dec 23, 2016·Journal of Medicinal Chemistry·Joyanta K ModakAnna Roujeinikova

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