1999
Trafficking of Endothelial Nitric-oxide Synthase in Living Cells QUANTITATIVE EVIDENCE SUPPORTING THE ROLE OF PALMITOYLATION AS A KINETIC TRAPPING MECHANISM LIMITING MEMBRANE DIFFUSION*
Sowa G, Liu J, Papapetropoulos A, Rex-Haffner M, Hughes T, Sessa W. Trafficking of Endothelial Nitric-oxide Synthase in Living Cells QUANTITATIVE EVIDENCE SUPPORTING THE ROLE OF PALMITOYLATION AS A KINETIC TRAPPING MECHANISM LIMITING MEMBRANE DIFFUSION*. Journal Of Biological Chemistry 1999, 274: 22524-22531. PMID: 10428829, DOI: 10.1074/jbc.274.32.22524.Peer-Reviewed Original ResearchConceptsPlasma membraneENOS-GFPFluorescent protein fusion constructsProtein-protein interactionsRole of palmitoylationProtein fusion constructsLipid bilayersRate of traffickingEndothelial cell line ECV304Endothelial nitric oxide synthasePalmitoylation stateCellular domainsFusion constructsPerinuclear regionLiving cellsProtein diffusionFluorescence recoveryRegulation of eNOSMembrane markersPalmitoylationMutantsGolgiTraffickingMembrane diffusionEndothelial cells
1997
Dissecting the Interaction between Nitric Oxide Synthase (NOS) and Caveolin FUNCTIONAL SIGNIFICANCE OF THE NOS CAVEOLIN BINDING DOMAININ VIVO *
Garcı́a-Cardeña G, Martasek P, Masters B, Skidd P, Couet J, Li S, Lisanti M, Sessa W. Dissecting the Interaction between Nitric Oxide Synthase (NOS) and Caveolin FUNCTIONAL SIGNIFICANCE OF THE NOS CAVEOLIN BINDING DOMAININ VIVO *. Journal Of Biological Chemistry 1997, 272: 25437-25440. PMID: 9325253, DOI: 10.1074/jbc.272.41.25437.Peer-Reviewed Original ResearchConceptsCaveolin-1Peripheral membrane proteinsInteraction of eNOSC-terminal tailAmino acids 310Direct interactionCo-transfection experimentsSite-directed mutagenesisNovel functional roleEndothelial nitric oxide synthaseMolecular chaperonesCytoplasmic domainCaveolin isoformsDeletion mutantsMammalian cellsEndothelial cell lysatesGlutathione S-transferaseMembrane proteinsCaveolin-2Coat proteinNegative regulationCaveolin-3Endothelial cellsDirect bindingGolgi region
1996
Characterization of Bovine Endothelial Nitric Oxide Synthase Expressed inE. coli
Martasek P, Liu Q, Liu J, Roman L, Gross S, Sessa W, Masters B. Characterization of Bovine Endothelial Nitric Oxide Synthase Expressed inE. coli. Biochemical And Biophysical Research Communications 1996, 219: 359-365. PMID: 8604992, DOI: 10.1006/bbrc.1996.0238.Peer-Reviewed Original ResearchConceptsE. coliBaculovirus expression systemBovine endothelial nitric oxide synthaseSDS/PAGEHEK-293 cellsChaperonin GroELHeterologous expressionRecombinant enzymeExpression systemExpression vectorActive enzymeFormation assaysEnzymatic activityColiEnzymeTissue sourcesEndothelial constitutive nitric oxide synthaseProteinSingle bandSynthaseNitric oxide synthaseAbsorbance maximumEndothelial nitric oxide synthase
1995
Functional Analysis of the Human Endothelial Nitric Oxide Synthase Promoter SP1 AND GATA FACTORS ARE NECESSARY FOR BASAL TRANSCRIPTION IN ENDOTHELIAL CELLS (∗)
Zhang R, Min W, Sessa W. Functional Analysis of the Human Endothelial Nitric Oxide Synthase Promoter SP1 AND GATA FACTORS ARE NECESSARY FOR BASAL TRANSCRIPTION IN ENDOTHELIAL CELLS (∗). Journal Of Biological Chemistry 1995, 270: 15320-15326. PMID: 7541039, DOI: 10.1074/jbc.270.25.15320.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid OxidoreductasesAnimalsAortaBase SequenceBinding SitesCattleDNA PrimersDNA-Binding ProteinsEndothelium, VascularGATA2 Transcription FactorGene ExpressionHumansLuciferasesMolecular Sequence DataMutagenesis, Site-DirectedNitric Oxide SynthasePolymerase Chain ReactionPromoter Regions, GeneticRecombinant ProteinsRestriction MappingSp1 Transcription FactorTranscription FactorsTranscription, GeneticTransfectionConceptsBovine aortic endothelial cellsPromoter activitySp1 sitesBasal transcriptionGATA siteFurther deletionNuclear extractsENOS promoter activityAntibody supershift analysisDNA-protein complexesLuciferase activityENOS promoterLuc constructGel shift analysisSp-1 sitesBasal promoter activityLuciferase reporter geneSynthase gene expressionSp1 mutantGATA factorsNuclear proteinsGATA-2Mutant constructsReporter geneGel shift