2024
Mucosal sugars delineate pyrazine vs pyrazinone autoinducer signaling in Klebsiella oxytoca
Hamchand R, Wang K, Song D, Palm N, Crawford J. Mucosal sugars delineate pyrazine vs pyrazinone autoinducer signaling in Klebsiella oxytoca. Nature Communications 2024, 15: 8902. PMID: 39406708, PMCID: PMC11480411, DOI: 10.1038/s41467-024-53185-6.Peer-Reviewed Original ResearchConceptsK. oxytocaGeneral carbohydrate metabolismVirulence factor productionPLP-dependent enzymesAssociated with gutEnterobactin biosynthesisAutoinducer signalBacterial virulenceKlebsiella oxytocaSpecific carbohydratesHost immune responseCarbohydrate metabolismAutoinducerMolecular signalsVirulenceHistamine receptor H4BiosynthesisHost signalAcquisition responsesProtease inhibitorsPathwayHostLung pathologyLung isolationImmune response
2021
Escherichia coli-Derived γ‑Lactams and Structurally Related Metabolites Are Produced at the Intersection of Colibactin and Fatty Acid Biosynthesis
Kim CS, Turocy T, Moon G, Shine EE, Crawford JM. Escherichia coli-Derived γ‑Lactams and Structurally Related Metabolites Are Produced at the Intersection of Colibactin and Fatty Acid Biosynthesis. Organic Letters 2021, 23: 6895-6899. PMID: 34406772, PMCID: PMC10577019, DOI: 10.1021/acs.orglett.1c02461.Peer-Reviewed Original ResearchConceptsFatty acid biosynthesisAcid biosynthesisHybrid polyketide-nonribosomal peptideNuclear magnetic resonance spectroscopyΓ-lactam derivativesUnknown biological activityBiosynthetic logicMagnetic resonance spectroscopyCancer initiationColorectal cancer initiationStereochemical analysisΓ-lactamsColibactinResonance spectroscopyDiverse collectionBiosynthesisBiological activityAbundant metabolitesPathwayRelated metabolitesRelated structuresMetabolitesLociProteinSpectroscopy
2020
Making and Breaking Leupeptin Protease Inhibitors in Pathogenic Gammaproteobacteria
Li J, Oh J, Kienesberger S, Kim NY, Clarke DJ, Zechner EL, Crawford J. Making and Breaking Leupeptin Protease Inhibitors in Pathogenic Gammaproteobacteria. Angewandte Chemie International Edition 2020, 59: 17872-17880. PMID: 32609431, DOI: 10.1002/anie.202005506.Peer-Reviewed Original ResearchConceptsBacterial small moleculesTripeptide aldehyde protease inhibitorsHeterologous expression analysisProtease inhibitorsColonization phenotypesPhotorhabdus speciesHypothetical proteinsNematode hostsPathogenic GammaproteobacteriaExpression analysisBacterial productionAccessory enzymesGenetic distributionProteolytic degradationSmall moleculesEnzymeLeupeptinKlebsiella oxytocaInhibitorsGammaproteobacteriaLigasesXenorhabdusPhotorhabdusBiosynthesisSpeciesMaking and Breaking Leupeptin Protease Inhibitors in Pathogenic Gammaproteobacteria
Li J, Oh J, Kienesberger S, Kim N, Clarke D, Zechner E, Crawford J. Making and Breaking Leupeptin Protease Inhibitors in Pathogenic Gammaproteobacteria. Angewandte Chemie 2020, 132: 18028-18036. DOI: 10.1002/ange.202005506.Peer-Reviewed Original ResearchBacterial small moleculesTripeptide aldehyde protease inhibitorsHeterologous expression analysisProtease inhibitorsPhotorhabdus speciesColonization phenotypesHypothetical proteinsNematode hostsPathogenic GammaproteobacteriaExpression analysisBacterial productionAccessory enzymesGenetic distributionProteolytic degradationSmall moleculesEnzymeLeupeptinKlebsiella oxytocaInhibitorsLigasesXenorhabdusGammaproteobacteriaPhotorhabdusBiosynthesisSpeciesCharacterization of Autoinducer‑3 Structure and Biosynthesis in E. coli
Kim CS, Gatsios A, Cuesta S, Lam YC, Wei Z, Chen H, Russell RM, Shine EE, Wang R, Wyche TP, Piizzi G, Flavell RA, Palm NW, Sperandio V, Crawford JM. Characterization of Autoinducer‑3 Structure and Biosynthesis in E. coli. ACS Central Science 2020, 6: 197-206. PMID: 32123737, PMCID: PMC7047286, DOI: 10.1021/acscentsci.9b01076.Peer-Reviewed Original ResearchAutoinducer-3Population-level phenotypesDiverse biological rolesGram-positive bacterial pathogensPrimary human tissuesModel organismsQuorum sensingMolecular foundationBiological roleCommon inhabitantsVariety of GramHuman microbiotaVirulence genesBacterial pathogensE. coliBiochemical originBiosynthesisHuman tissuesUnknown structureSynthetase reactionGenesOrganismsBiologyMetabolitesEnterohemorrhagic
2018
Characterization of Natural Colibactin–Nucleobase Adducts by Tandem Mass Spectrometry and Isotopic Labeling. Support for DNA Alkylation by Cyclopropane Ring Opening
Xue M, Shine E, Wang W, Crawford JM, Herzon SB. Characterization of Natural Colibactin–Nucleobase Adducts by Tandem Mass Spectrometry and Isotopic Labeling. Support for DNA Alkylation by Cyclopropane Ring Opening. Biochemistry 2018, 57: 6391-6394. PMID: 30365310, PMCID: PMC6997931, DOI: 10.1021/acs.biochem.8b01023.Peer-Reviewed Original ResearchConceptsGenotoxic secondary metabolitesMethionine auxotrophic strainsE. coliGut commensal Escherichia coliClb gene clusterColibactin biosynthesisElectrophilic CyclopropanesGene clusterDNA alkylationPrevious biosynthetic studiesSecondary metabolitesAuxotrophic strainsEscherichia coliCommensal Escherichia coliBiosynthetic studiesFirst identificationBiosynthesisColiColibactinPUC19 DNAIsotopic labelingNucleotidesDNATandem mass spectrometryCertain strains
2017
ClbS Is a Cyclopropane Hydrolase That Confers Colibactin Resistance
Tripathi P, Shine EE, Healy AR, Kim CS, Herzon SB, Bruner SD, Crawford JM. ClbS Is a Cyclopropane Hydrolase That Confers Colibactin Resistance. Journal Of The American Chemical Society 2017, 139: 17719-17722. PMID: 29112397, PMCID: PMC6202678, DOI: 10.1021/jacs.7b09971.Peer-Reviewed Original ResearchConceptsGene productsBiosynthetic gene clusterSpecific mechanistic roleMolecular functionsGene clusterResidue mutantsHost bacteriaCancer formationColorectal cancer formationEscherichia coliCommensal Escherichia coliColibactinMechanistic roleHydrolase activityPrecolibactinsX-ray structureMolecular-level viewShare similaritiesElectrophilic CyclopropanesBacterial viabilityBacteriaHydrolaseGenotoxic effectsMutantsBiosynthesisDomain-Targeted Metabolomics Delineates the Heterocycle Assembly Steps of Colibactin Biosynthesis
Trautman EP, Healy AR, Shine EE, Herzon SB, Crawford JM. Domain-Targeted Metabolomics Delineates the Heterocycle Assembly Steps of Colibactin Biosynthesis. Journal Of The American Chemical Society 2017, 139: 4195-4201. PMID: 28240912, PMCID: PMC5831107, DOI: 10.1021/jacs.7b00659.Peer-Reviewed Original ResearchConceptsNonribosomal peptide synthetasesPolyketide synthasesModular polyketide synthasesSite-directed mutagenesisCombination of genesCellular metabolic levelsColibactin biosynthesisMultidomain proteinsProtein domainsPeptide synthetasesCatalytic domainControl pathwaysProtein biochemicalPathway analysisCertain Escherichia coliComplete deletionHybrid pathwayEscherichia coliFunctional readoutBiosynthesisCatalytic mechanismCellular levelMetabolic levelPathwayGenes
2015
The colibactin warhead crosslinks DNA
Vizcaino MI, Crawford JM. The colibactin warhead crosslinks DNA. Nature Chemistry 2015, 7: 411-417. PMID: 25901819, PMCID: PMC4499846, DOI: 10.1038/nchem.2221.Peer-Reviewed Original ResearchConceptsDNA-damaging activityHost-microbe interactionsUncharacterized familyColibactin biosynthesisSmall moleculesHybrid pathwayCancer formationMode of actionDuplex DNACrosslinks DNAMicrobial metabolitesUnexpected modelHuman microbiotaSelect strainsMetabolomic analysisDNAPathwayEscherichia coli presentHuman colonLabeling studiesDisease statesDirect experimental evidenceHuman healthBiosynthesisColibactin
2012
Small molecule perimeter defense in entomopathogenic bacteria
Crawford JM, Portmann C, Zhang X, Roeffaers MB, Clardy J. Small molecule perimeter defense in entomopathogenic bacteria. Proceedings Of The National Academy Of Sciences Of The United States Of America 2012, 109: 10821-10826. PMID: 22711807, PMCID: PMC3390839, DOI: 10.1073/pnas.1201160109.Peer-Reviewed Original ResearchConceptsBacterial cell surfaceConfocal fluorescence microscopyCell surfacePhysical associationHuman pathogen Vibrio choleraeInsect innate immune systemBiochemical assaysInnate immune systemPathogen Vibrio choleraeFluorescence microscopyPrecursor-directed biosynthesisE. coli cellsInsect pathogensPathway mutantsEntomopathogenic bacteriaHeterologous expressionXenorhabdus nematophilaColi cellsEscherichia coliVibrio choleraeClass of moleculesBioorthogonal probesEffective inhibitor concentrationBiosynthesisHost defense
2010
Not just passing through
Crawford JM, Clardy J. Not just passing through. Nature Chemistry 2010, 2: 805-807. PMID: 20861892, DOI: 10.1038/nchem.848.Peer-Reviewed Original ResearchAbsence of the aflatoxin biosynthesis gene, norA, allows accumulation of deoxyaflatoxin B1 in Aspergillus flavus cultures
Ehrlich KC, Chang P, Scharfenstein LL, Cary JW, Crawford JM, Townsend CA. Absence of the aflatoxin biosynthesis gene, norA, allows accumulation of deoxyaflatoxin B1 in Aspergillus flavus cultures. FEMS Microbiology Letters 2010, 305: 65-70. PMID: 20158523, PMCID: PMC2891446, DOI: 10.1111/j.1574-6968.2010.01914.x.Peer-Reviewed Original ResearchConceptsBiosynthetic gene clusterAflatoxin biosynthesis genesCytochrome P450 monooxygenaseAflatoxin biosynthetic gene clusterWild-type A. flavusBiosynthesis genesGene clusterP450 monooxygenaseAflatoxin biosynthesisCarcinogenic aflatoxinsAccumulation of aflatoxinBiosynthesisA. flavusMinor toxinsO-methylsterigmatocystinAspergillus speciesFinal stepAspergillus flavusNew metabolitesQuantities of aflatoxinElevated quantitiesMutantsAccumulationFlavusGenes
2008
Acyl‐Carrier Protein–Phosphopantetheinyltransferase Partnerships in Fungal Fatty Acid Synthases
Crawford JM, Vagstad AL, Ehrlich KC, Udwary DW, Townsend CA. Acyl‐Carrier Protein–Phosphopantetheinyltransferase Partnerships in Fungal Fatty Acid Synthases. ChemBioChem 2008, 9: 1559-1563. PMID: 18551496, PMCID: PMC3189688, DOI: 10.1002/cbic.200700659.Peer-Reviewed Original ResearchDeconstruction of Iterative Multidomain Polyketide Synthase Function
Crawford J, Thomas P, Scheerer J, Vagstad A, Kelleher N, Townsend C. Deconstruction of Iterative Multidomain Polyketide Synthase Function. Science 2008, 320: 243-246. PMID: 18403714, PMCID: PMC2480491, DOI: 10.1126/science.1154711.Peer-Reviewed Original ResearchConceptsIterative polyketide synthasesIndividual catalytic domainsProduct template (PT) domainsPolyketide synthasesStarter unitCatalytic domainAromatic polyketidesBiosynthetic enzymesTemplate domainSynthase functionCarcinogen aflatoxin B1PksAMechanistic featuresEnzymePt domainsKetosynthaseDomainPolyketidesBiosynthesisSynthasesCascadeReconstitutionAccumulationFamily