The tricarboxylic acid cycle of Helicobacter pylori.
The composition and properties of the tricarboxylic acid cycle of the microaerophilic human pathogen Helicobacter pylori were investigated in situ and in cell extracts using [1H]- and [13C]-NMR spectroscopy and spectrophotometry. NMR spectroscopy assays enabled highly specific measurements of some enzyme activities, previously not possible using spectrophotometry, in in situ studies with H. pylori, thus providing the first accurate picture of the complete tricarboxylic acid cycle of the bacterium. The presence, cellular location and kinetic parameters of citrate synthase, aconitase, isocitrate dehydrogenase, alpha-ketoglutarate oxidase, fumarate reductase, fumarase, malate dehydrogenase, and malate synthase activities in H. pylori are described. The absence of other enzyme activities of the cycle, including alpha-ketoglutarate dehydrogenase, succinyl-CoA synthetase, and succinate dehydrogenase also are shown. The H. pylori tricarboxylic acid cycle appears to be a noncyclic, branched pathway, characteristic of anaerobic metabolism, directed towards the production of succinate in the reductive dicarboxylic acid branch and alpha-ketoglutarate in the oxidative tricarboxylic acid branch. Both branches were metabolically linked by the presence of alpha-ketoglutarate oxidase activity. Under the growth conditions employed, H. pylori did not possess an operational glyoxylate bypass, owing to the absence of isocitrate lyase activity; nor a gamma-aminobutyrate shunt, owing to the absence of both gamma-aminobutyrate transaminase and succinic semialdehyde dehydrogenase activities. The catalytic and regulatory properties of the H. pylori tricarboxylic acid cycle enzymes are discussed by comparing their amino acid sequences with those of other, more extensively studied enzymes.
Pitson
S M
SM
School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, Australia.
Mendz
G L
GL
Srinivasan
S
S
Hazell
S L
SL
D000494:Allosteric Regulation; D002952:Citric Acid Cycle / Q000502:physiology*; D003065:Coenzyme A / Q000378:metabolism; D016680:Genome, Bacterial; D006038:Glyoxylates / Q000378:metabolism; D016480:Helicobacter pylori / Q000201:enzymology*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D019802:Succinic Acid / Q000378:metabolism
D006038:Glyoxylates; D019802:Succinic Acid; C031150:glyoxylic acid; D003065:Coenzyme A