title: A new regulatory principle for in vivo biochemistry: pleiotropic low affinity regulation by the adenine nucleotides--illustrated for the glycolytic enzymes of Saccharomyces cerevisiae.
abstract: Enzymology tends to focus on highly specific effects of substrates, allosteric modifiers, and products occurring at low concentrations, because these are most informative about the enzyme's catalytic mechanism. We hypothesized that at relatively high in vivo concentrations, important molecular monitors of the state of living cells, such as ATP, affect multiple enzymes of the former and that these interactions have gone unnoticed in enzymology. We test this hypothesis in terms of the effect that ATP, ADP, and AMP might have on the major free-energy delivering pathway of the yeast Saccharomyces cerevisiae. Assaying cell-free extracts, we collected a comprehensive set of quantitative kinetic data concerning the enzymes of the glycolytic and the ethanol fermentation pathways. We determined systematically the extent to which the enzyme activities depend on the concentrations of the adenine nucleotides. We found that the effects of the adenine nucleotides on enzymes catalysing reactions in which they are not directly involved as substrate or product, are substantial. This includes effects on the Michaelis-Menten constants, adding new perspective on these, 100 years after their introduction.
authors.affiliation: Department of Molecular Cell Physiology, Faculty of Earth and Life Sciences, VU University Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
authors.forename: Femke I C
authors.initials: FI
authors.lastname: Mensonides
authors.forename: Barbara M
authors.initials: BM
authors.lastname: Bakker
authors.forename: Frederic
authors.initials: F
authors.lastname: Cremazy
authors.forename: Hanan L
authors.initials: HL
authors.lastname: Messiha
authors.forename: Pedro
authors.initials: P
authors.lastname: Mendes
authors.forename: Fred C
authors.initials: FC
authors.lastname: Boogerd
authors.forename: Hans V
authors.initials: HV
authors.lastname: Westerhoff
mesh_terms: D000227:Adenine Nucleotides / Q000737:chemistry* / Q000502:physiology; D000494:Allosteric Regulation; D005285:Fermentation; D006019:Glycolysis; D007700:Kinetics; D008954:Models, Biological; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D013347:Subcellular Fractions / Q000201:enzymology; D013816:Thermodynamics
chemical_list: D000227:Adenine Nucleotides; D029701:Saccharomyces cerevisiae Proteins
keywords: 2-phosphoglyceric acid; 2PGA; 3-phosphoglycerate kinase (EC 2.7.2.3); 3-phosphoglyceric acid; 3PGA; ADH; ALD; ATP; ENO; Energetics; Enzyme kinetics; EtOH; F16BP; F26BP; F6P; G3PDH; G6P; G6PDH; GAP; GAPDH; Glc; HXK; LDH; PDC; PEP; PFK; PGI; PGK; PGM; PYK; Pyr; Systems Biology; TPI; Yeast glycolysis model; alcohol dehydrogenase (EC 1.1.1.1); enolase (EC 4.2.1.11); ethanol; fructose 1,6-bisphosphate; fructose 1,6-bisphosphate aldolase (EC 4.1.2.13); fructose 2,6-bisphosphate; fructose 6-phosphate; glucose; glucose 6-phosphate; glucose-6-phosphate dehydrogenase (EC 1.1.1.49); glyceraldehyde 3-phosphate; glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12); glycerol 3-phosphate dehydrogenase (EC 1.1.1.8); hexokinase (EC 2.7.1.1); lactate dehydrogenase (EC 1.1.1.27); phosphoenolpyruvate; phosphofructokinase (EC 2.7.1.11); phosphoglucose isomerase (EC 5.3.1.9); phosphoglycerate mutase (EC 5.4.2.1); pyruvate; pyruvate decarboxylase (EC 4.1.1.1); pyruvate kinase (EC 2.7.1.40); triosephosphate isomerase (EC 5.3.1.1)