title: Systematic identification of allosteric protein-metabolite interactions that control enzyme activity in vivo.
abstract: Recent data suggest that the majority of proteins bind specific metabolites and that such interactions are relevant to metabolic and gene regulation. However, there are no methods to systematically identify functional allosteric protein-metabolite interactions. Here we present an experimental and computational approach for using dynamic metabolite data to discover allosteric regulation that is relevant in vivo. By switching the culture conditions of Escherichia coli every 30 s between medium containing either pyruvate or (13)C-labeled fructose or glucose, we measured the reversal of flux through glycolysis pathways and observed rapid changes in metabolite concentration. We fit these data to a kinetic model of glycolysis and systematically tested the consequences of 126 putative allosteric interactions on metabolite dynamics. We identified allosteric interactions that govern the reversible switch between gluconeogenesis and glycolysis, including one by which pyruvate activates fructose-1,6-bisphosphatase. Thus, from large sets of putative allosteric interactions, our approach can identify the most likely ones and provide hypotheses about their function.
authors: Institute of Molecular System Biology, ETH Zurich, Zurich, Switzerland.
authors: Hannes
authors: H
authors: Link
authors: Karl
authors: K
authors: Kochanowski
authors: Uwe
authors: U
authors: Sauer
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D001426:Bacterial Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology*; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005943:Gluconeogenesis / Q000187:drug effects; D006019:Glycolysis / Q000187:drug effects; D006601:Hexoses / Q000494:pharmacology; D055442:Metabolome / Q000187:drug effects; D019289:Pyruvic Acid / Q000494:pharmacology; D015203:Reproducibility of Results; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D001426:Bacterial Proteins; D006601:Hexoses; D019289:Pyruvic Acid; D006597:Fructose-Bisphosphatase