From genome to enzyme: analysis of key glycolytic and oxidative pentose-phosphate pathway enzymes in the cyanobacterium Synechocystis sp. PCC 6803.
Activities of glucokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, phosphoglucose isomerase, phosphofructokinase (PFK), enolase, pyruvate kinase (PK) and phosphoenolpyruvate (PEP) carboxylase were determined in extracts of photoautotrophic, mixotrophic, and heterotrophic cultures of Synechocystis sp. PCC 6803. Annotated genomes of Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120 were analyzed for the respective predicted physical properties of each enzyme investigated here. Enzymatic activity was largely unaffected by nutritional mode, with the exception of glucokinase and PK whose activities were significantly elevated in heterotrophic cultures of Synechocystis sp. PCC 6803. PFK activity was insensitive to bacterial PFK-A (allosteric) effectors such as PEP, implying that Synechocystis PFK should be classified as a PFK-B (non-allosteric). Immunoblot and kinetic studies indicated that irrespective of nutritional mode, the Synechocystis PK corresponds to a PK-A (AMP activated) rather than PK-F (fructose-1,6-bisphosphate activated).
Knowles
Vicki L
VL
Department of Biology, Queen's University, Kingston, Ontario, K7L 3N6 Canada.
Plaxton
William C
WC
D017033:Anabaena / Q000201:enzymology / Q000235:genetics; D000458:Cyanobacteria / Q000201:enzymology* / Q000235:genetics; D016680:Genome, Bacterial; D005941:Glucokinase / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006019:Glycolysis; D010427:Pentose Phosphate Pathway; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism; D025221:Phosphofructokinases / Q000378:metabolism; D010734:Phosphogluconate Dehydrogenase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism
D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D025221:Phosphofructokinases; D005941:Glucokinase; D011770:Pyruvate Kinase; D010730:Phosphoenolpyruvate Carboxylase; D010751:Phosphopyruvate Hydratase; D005956:Glucose-6-Phosphate Isomerase