Characterization of the Helicobacter pylori urease and purification of its subunits.
Helicobacter pylori (formerly Campylobacter pylori) is the causative agent of gastritis in man. Helicobacter pylori cells contain a large amount of an extremely active urease (E.C.3.5.1.5). This enzyme is suspected to be a virulence factor since the ammonium ion produced from urea may be responsible for tissue injury and/or survival of H. pylori in the gastric environment. Helicobacter pylori urease, native relative molecular mass approximately 600,000, was purified by agarose gel filtration and ion exchange chromatography. DEAE-purified urease is highly active and has a Km of 0.48 mM for urea. The enzyme has a pI of 5.93 and is active from pH 4.0 to 10.0, with an optimum at pH 8.0. The purified urease contains nickel and is composed of two protein subunits, with relative molecular masses of 66,000 and 31,000. The subunits were separated and purified and the first 30 N-terminal amino acid residues were determined. A remarkably close relationship was found between both H. pylori urease subunits and jack bean (Canavalia ensiformis) urease, the subunit of which is a single 840 amino acid polypeptide. This subunit is also largely identical to the high molecular mass subunits of the ureases of Klebsiella aerogenes and Proteus mirabilis, evidence that these four ureases are derived from a common ancestral protein.
Evans
D J
DJ
Department of Medicine, Baylor College of Medicine, Houston, Texas.
Evans
D G
DG
Kirkpatrick
S S
SS
Graham
D Y
DY
D000595:Amino Acid Sequence; D002845:Chromatography; D007887:Fabaceae / Q000201:enzymology; D016480:Helicobacter pylori / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009532:Nickel / Q000032:analysis; D010946:Plants, Medicinal; D011513:Proteus mirabilis / Q000201:enzymology; D014510:Urease / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D046911:Macromolecular Substances; D009532:Nickel; D014510:Urease