Purification of Helicobacter pylori superoxide dismutase and cloning and sequencing of the gene.
The superoxide dismutase (SOD) of Helicobacter pylori, a pathogenic bacterium which colonizes the gastric mucosa, evoking a marked inflammatory response, was purified and characterized, and the N-terminal amino acid sequence was determined. The enzyme consists of two identical subunits each with an apparent molecular weight of 24,000. Analysis of the primary structure and inhibition studies revealed that H. pylori possesses a typical procaryotic iron-containing enzyme. No other isoenzymes could be detected. Indirect gold immunostaining of H. pylori SOD with a polyclonal antibody directed against the iron-containing SOD of Escherichia coli showed a surface-associated localization of the enzyme. The H. pylori SOD gene was cloned by functional complementation of a SOD-deficient E. coli mutant. Sequencing and alignment revealed striking homology to the following facultative intracellular human pathogens: Listeria ivanovii, Listeria monocytogenes, Coxiella burnetti, Porphyromonas gingivalis, Legionella pneumophila, and Entamoeba histolytica. An open reading frame of 642 bp encoding 214 amino acids was determined. There was no leader sequence detectable. Cloning of the H. pylori SOD gene is one of the prerequisites to investigation of its pathophysiological role in the defense against antimicrobial mechanisms of polymorphonuclear granulocytes.
Spiegelhalder
C
C
Institute for Medical Microbiology and Hygiene, Freiburg, Germany.
Gerstenecker
B
B
Kersten
A
A
Schiltz
E
E
Kist
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D016481:Helicobacter Infections / Q000201:enzymology; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics* / Q000648:ultrastructure; D006801:Humans; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism
D004269:DNA, Bacterial; D013482:Superoxide Dismutase