1999
The Catalytic Mechanism of EPSP Synthase Revisited †
Lewis J, Johnson K, Anderson K. The Catalytic Mechanism of EPSP Synthase Revisited †. Biochemistry 1999, 38: 7372-7379. PMID: 10353849, DOI: 10.1021/bi9830258.Peer-Reviewed Original ResearchMeSH Keywords3-Phosphoshikimate 1-CarboxyvinyltransferaseAlkyl and Aryl TransferasesAmino Acid SubstitutionBinding SitesCatalysisChromatography, High Pressure LiquidEscherichia coliFreezingKineticsMutagenesis, Site-DirectedNuclear Magnetic Resonance, BiomolecularPhosphoenolpyruvateProtonsSubstrate SpecificityConceptsEPSP synthaseEnzyme intermediateKinetic competenceSingle-turnover experimentsSubstrate to productSolid-state NMRSolid-state NMR studiesEnzyme assaysEnzyme reaction pathwaySDS-PAGECatalytic mechanismDegrees CSpeciesEnzymeIntermediate speciesNMR studiesSide productsCharacterized reaction productsSample preparationDisappearance of substrateSynthaseReaction productsFormation of productsBreakdown productsReaction pathways
1997
Detection and Identification of Transient Enzyme Intermediates Using Rapid Mixing, Pulsed-Flow Electrospray Mass Spectrometry †
Paiva A, Tilton R, Crooks G, Huang L, Anderson K. Detection and Identification of Transient Enzyme Intermediates Using Rapid Mixing, Pulsed-Flow Electrospray Mass Spectrometry †. Biochemistry 1997, 36: 15472-15476. PMID: 9398276, DOI: 10.1021/bi971883i.Peer-Reviewed Original ResearchMeSH Keywords3-Phosphoshikimate 1-CarboxyvinyltransferaseAlkyl and Aryl TransferasesCatalysisMass SpectrometryConceptsTetrahedral intermediateElectrospray ionization ion trap mass spectrometerIon trap mass spectrometerNegative ion mass spectraElectrospray ionization mass spectrometryCollision-induced dissociationEnzyme intermediateIon mass spectraTrap mass spectrometerIonization mass spectrometryEnzyme reaction intermediatesElectrospray ionizationDaughter ionsSubsecond time scaleEnzyme active siteReaction intermediatesAtomic mass unitsMass spectraMass spectrometerChemical quench studiesQuenching studiesMass spectrometryRapid mixing deviceQuenching methodActive site
1995
Reevaluating glyphosate as a transition-state inhibitor of EPSP synthase: identification of an EPSP synthase.EPSP.glyphosate ternary complex.
Sammons R, Gruys K, Anderson K, Johnson K, Sikorski J. Reevaluating glyphosate as a transition-state inhibitor of EPSP synthase: identification of an EPSP synthase.EPSP.glyphosate ternary complex. Biochemistry 1995, 34: 6433-40. PMID: 7756274, DOI: 10.1021/bi00019a024.Peer-Reviewed Original ResearchMeSH Keywords3-Phosphoshikimate 1-CarboxyvinyltransferaseAlkyl and Aryl TransferasesGlycineKineticsMacromolecular SubstancesProtein BindingRecombinant ProteinsTransferasesConceptsEPSP synthaseTernary complexShikimate 3-phosphateSteady-state kineticsEnzyme active siteTransition-state analogSubstrate turnoverSynthase reactionTransition-state inhibitorsEnzymeAssociated with PEPUncompetitive inhibitorBinding resultsSynthaseActive siteFluorescence titration experimentsShikimateOxonium ionsTurnoverInteraction of glyphosateTitration experiments
1994
Detection and characterization of a phospholactoyl-enzyme adduct in the reaction catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase, MurZ.
Brown E, Marquardt J, Lee J, Walsh C, Anderson K. Detection and characterization of a phospholactoyl-enzyme adduct in the reaction catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase, MurZ. Biochemistry 1994, 33: 10638-45. PMID: 8075064, DOI: 10.1021/bi00201a010.Peer-Reviewed Original ResearchMeSH KeywordsAlkyl and Aryl TransferasesBacterial ProteinsBinding SitesCatalysisEscherichia coliKineticsMagnetic Resonance SpectroscopyPhosphoenolpyruvateTransferasesConceptsUDP-N-acetylglucosamineUDP-GlcNAcAbsence of UDP-GlcNAcChemical quench analysisPresence of UDP-GlcNAcSingle-turnover conditionsBinding constantsPeptidoglycan biosynthesisSolution NMRC-2Enzyme nucleophilePeptide of molecular weightStoichiometric labelingConsistent with catalysisRemoval of small moleculesE. coliAdductsSmall moleculesMurZEnzyme adductNon-covalentlySDS-PAGEM ureaLabeled peptidesEnzyme
1990
"Kinetic competence" of the 5-enolpyruvoylshikimate-3-phosphate synthase tetrahedral intermediate.
Anderson K, Johnson K. "Kinetic competence" of the 5-enolpyruvoylshikimate-3-phosphate synthase tetrahedral intermediate. Journal Of Biological Chemistry 1990, 265: 5567-5572. PMID: 2180929, DOI: 10.1016/s0021-9258(19)39398-6.Peer-Reviewed Original ResearchObservation by 13C NMR of the EPSP synthase tetrahedral intermediate bound to the enzyme active site.
Anderson K, Sammons R, Leo G, Sikorski J, Benesi A, Johnson K. Observation by 13C NMR of the EPSP synthase tetrahedral intermediate bound to the enzyme active site. Biochemistry 1990, 29: 1460-5. PMID: 2334707, DOI: 10.1021/bi00458a017.Peer-Reviewed Original ResearchConceptsEnzyme active siteTetrahedral intermediateFormation of pyruvateActive siteEnzyme sitesComparison of quenchingReaction of enzymeTime of incubationTetrahedral centerCompound giving riseReaction pathwaysEnzymatic hydrolysisPeak assignmentsEnzymeNMR experimentsTernary complexNMR measurementsSide productsRate of formationSpectroscopic probesLong time of incubationNMRSpeciesTriethylamineCovalent adducts
1988
A tetrahedral intermediate in the EPSP synthase reaction observed by rapid quench kinetics.
Anderson K, Sikorski J, Johnson K. A tetrahedral intermediate in the EPSP synthase reaction observed by rapid quench kinetics. Biochemistry 1988, 27: 7395-406. PMID: 3061457, DOI: 10.1021/bi00419a034.Peer-Reviewed Original ResearchConceptsPhosphoenol pyruvateBurst of product formationPre-steady-state burstQuantitation of reaction productsTransient-state kinetic analysisEnzyme-bound intermediateShikimate 3-phosphateSingle turnover experimentsPre-steady-stateSubstrate trapping experimentsRelease of substratesEquilibrium constantsSynthase reactionExcess enzymeBinding rateAbsence of phosphatePyruvateReverse reactionEnzymeTurnover experimentsEnzymatic reactionsKinetic competenceEnzyme concentrationFormation of productsConcentration of phosphateEvaluation of 5-enolpyruvoylshikimate-3-phosphate synthase substrate and inhibitor binding by stopped-flow and equilibrium fluorescence measurements.
Anderson K, Sikorski J, Johnson K. Evaluation of 5-enolpyruvoylshikimate-3-phosphate synthase substrate and inhibitor binding by stopped-flow and equilibrium fluorescence measurements. Biochemistry 1988, 27: 1604-10. PMID: 3284585, DOI: 10.1021/bi00405a032.Peer-Reviewed Original ResearchMeSH Keywords3-Phosphoshikimate 1-CarboxyvinyltransferaseAlkyl and Aryl TransferasesEscherichia coliKineticsModels, TheoreticalProtein BindingSpectrometry, FluorescenceThermodynamicsTransferasesConceptsBinding of substratesBinary complexShikimate 3-phosphateStopped-flow fluorescence methodsDissociation constantFree enzymeGlyphosate bindingS3P bindingInhibitor bindingProtein fluorescenceKinetics of bindingTernary complexEnzymeStopped-flowFluorescence measurementsBindingFluorescence titrationSaturating concentrationsS3PEquilibrium fluorescence measurements