Cloning and characterization of Helicobacter pylori succinyl CoA:acetoacetate CoA-transferase, a novel prokaryotic member of the CoA-transferase family.
Sequencing of a fragment of Helicobacter pylori genome led to the identification of two open reading frames showing striking homology with Coenzyme A (CoA) transferases, enzymes catalyzing the reversible transfer of CoA from one carboxylic acid to another. The genes were present in all H. pylori strains tested by polymerase chain reaction or slot blotting but not in Campylobacter jejuni. Genes for the putative A and B subunits of H. pylori CoA-transferase were introduced into the bacterial expression vector pKK223-3 and expressed in Escherichia coli JM105 cells. Amino acid sequence comparisons, combined with measurements of enzyme activities using different CoA donors and acceptors, identified the H. pylori CoA-transferase as a succinyl CoA:acetoacetate CoA-transferase. This activity was consistently observed in different H. pylori strains. Antibodies raised against either recombinant A or B subunits recognized two distinct subunits of Mr approximately 26,000 and 24, 000 that are both necessary for H. pylori CoA-transferase function. The lack of alpha-ketoglutarate dehydrogenase and of succinyl CoA synthetase activities indicates that the generation of succinyl CoA is not mediated by the tricarboxylic acid cycle in H. pylori. We postulate the existence of an alternative pathway where the CoA-transferase is essential for energy metabolism.
Corthésy-Theulaz
I E
IE
Division of Gastroenterology, Centre Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland. icorthes@ulys.unil.ch
Bergonzelli
G E
GE
Henry
H
H
Bachmann
D
D
Schorderet
D F
DF
Blum
A L
AL
Ornston
L N
LN
D000214:Acyl Coenzyme A / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D016123:Campylobacter jejuni / Q000201:enzymology; D003001:Cloning, Molecular; D015256:Coenzyme A-Transferases / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D015870:Gene Expression; D016480:Helicobacter pylori / Q000201:enzymology*; D006801:Humans; D008956:Models, Chemical; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction
D000214:Acyl Coenzyme A; D004269:DNA, Bacterial; C012046:succinyl-coenzyme A; D015256:Coenzyme A-Transferases; C021460:3-ketoacid CoA-transferase