Cloning and functional characterization of an enzyme from Helicobacter pylori that catalyzes two steps of the methylerythritol phosphate pathway for isoprenoid biosynthesis.
BACKGROUND
The methylerythritol phosphate pathway for isoprenoid biosynthesis is an attractive target for the design of new specific antibiotics for the treatment of gastrointestinal diseases associated with the presence of the bacterium Helicobacter pylori since this pathway which is essential to the bacterium is absent in humans.


RESULTS
This work reports the molecular cloning of one of the genes of the methylerythritol phosphate pathway form H. pylori (ispDF; HP_1440) its expression in Escherichia coli and the functional characterization of the recombinant enzyme. As shown by genetic complementation and in vitro functional assays the product of the ispDF gene form H. pylori is a bifunctional enzyme which can replace both CDP-methylerythritol synthase and methylerythritol cyclodiphosphate synthase from E. coli.


CONCLUSIONS
Designing inhibitors that affect at the same time both enzyme activities of the H. pylori bifunctional enzyme (i.e. by disrupting protein oligomerization) would result in more effective antibiotics which would be able to continue their action even if the bacterium acquired a resistance to another antibiotic directed against one of the individual activities.


CONCLUSIONS
The bifunctional enzyme would be an excellent target for the design of new, selective antibiotics for the treatment of H. pylori associated diseases.
Pérez-Gil
Jordi
J
Departament de Bioquimica i Biologia Molecular. Universitat de Barcelona, Avda Diagonal 645. 08028-Barcelona, Spain.
Bergua
María
M
Boronat
Albert
A
Imperial
Santiago
S
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D015195:Drug Design; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000627:therapeutic use; D004896:Erythritol / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D016481:Helicobacter Infections / Q000188:drug therapy / Q000201:enzymology* / Q000235:genetics; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics; D019761:Phosphorus-Oxygen Lyases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D013729:Terpenes; D019761:Phosphorus-Oxygen Lyases; D004896:Erythritol