2024
Identification of the potassium-binding site in serotonin transporter
Hellsberg E, Boytsov D, Chen Q, Niello M, Freissmuth M, Rudnick G, Zhang Y, Sandtner W, Forrest L. Identification of the potassium-binding site in serotonin transporter. Proceedings Of The National Academy Of Sciences Of The United States Of America 2024, 121: e2319384121. PMID: 38652746, PMCID: PMC11067047, DOI: 10.1073/pnas.2319384121.Peer-Reviewed Original ResearchConceptsSerotonin transporterSite-directed mutagenesis of residuesMutagenesis of residuesSite-directed mutagenesisHeterologous expression systemStudy of vesiclesNa2 siteClearance of serotoninPatch-clamp recordingsExpression systemBinding residuesSequential bindingMolecular dynamics simulationsBinding sitesPotassium binding siteSubstrate accumulationClamp recordingsVesiclesResiduesTurnover rateBindingStructural studiesChemical gradientsBinding configurationsSynaptic cleft
2023
Structure-based discovery of conformationally selective inhibitors of the serotonin transporter
Singh I, Seth A, Billesbølle C, Braz J, Rodriguiz R, Roy K, Bekele B, Craik V, Huang X, Boytsov D, Pogorelov V, Lak P, O'Donnell H, Sandtner W, Irwin J, Roth B, Basbaum A, Wetsel W, Manglik A, Shoichet B, Rudnick G. Structure-based discovery of conformationally selective inhibitors of the serotonin transporter. Cell 2023, 186: 2160-2175.e17. PMID: 37137306, PMCID: PMC10306110, DOI: 10.1016/j.cell.2023.04.010.Peer-Reviewed Original Research
2019
Serotonin transport in the 21st century
Rudnick G, Sandtner W. Serotonin transport in the 21st century. The Journal Of General Physiology 2019, 151: 1248-1264. PMID: 31570504, PMCID: PMC6829555, DOI: 10.1085/jgp.201812066.Peer-Reviewed Original Research
2016
Control of serotonin transporter phosphorylation by conformational state
Zhang YW, Turk BE, Rudnick G. Control of serotonin transporter phosphorylation by conformational state. Proceedings Of The National Academy Of Sciences Of The United States Of America 2016, 113: e2776-e2783. PMID: 27140629, PMCID: PMC4878475, DOI: 10.1073/pnas.1603282113.Peer-Reviewed Original ResearchConceptsTransmembrane helix 5Cytoplasmic permeation pathwaysOutward open conformationIntact rat basophilic leukemia cellsCGMP-dependent phosphorylationInhibition of phosphorylationTM5 helicesTransporter phosphorylationSERT regulationOutward openingCysteine residuesHelix 5Open conformationCytoplasmic endHuman SERTPhosphorylationPermeation pathwayConformational statesHeLa cellsRat basophilic leukemia cellsBasophilic leukemia cellsSERT activityExocytotic releaseLeukemia cellsMutations
2012
Cyclic GMP-dependent Stimulation of Serotonin Transport Does Not Involve Direct Transporter Phosphorylation by cGMP-dependent Protein Kinase*
Wong A, Zhang YW, Jeschke GR, Turk BE, Rudnick G. Cyclic GMP-dependent Stimulation of Serotonin Transport Does Not Involve Direct Transporter Phosphorylation by cGMP-dependent Protein Kinase*. Journal Of Biological Chemistry 2012, 287: 36051-36058. PMID: 22942288, PMCID: PMC3476273, DOI: 10.1074/jbc.m112.394726.Peer-Reviewed Original ResearchConceptsCGMP-dependent protein kinaseProtein kinaseATP analogUnidentified protein kinasesWild-type kinaseMitogen-activated protein kinaseP38 mitogen-activated protein kinasePhosphorylation site sequencePKG-dependent phosphorylationModel peptide substratesTransporter phosphorylationKinase cascadePhosphorylation sitesWT kinaseDirect substrateProtein substratesResidue mutantsSerotonin transporterPeptide library screeningSite sequenceP38 inhibitorLibrary screeningKinasePeptide substratesCultured cells
2009
Ligand Effects on Cross-linking Support a Conformational Mechanism for Serotonin Transport*
Tao Z, Zhang YW, Agyiri A, Rudnick G. Ligand Effects on Cross-linking Support a Conformational Mechanism for Serotonin Transport*. Journal Of Biological Chemistry 2009, 284: 33807-33814. PMID: 19837674, PMCID: PMC2797150, DOI: 10.1074/jbc.m109.071977.Peer-Reviewed Original ResearchConceptsN-terminal cyanogen bromide fragmentGamma-aminobutyric acid transporterCyanogen bromide fragmentsTransmembrane 1Cysteine residuesMutagenesis strategyAcid transportersConformational mechanismSerotonin transportCysteineCorresponding positionDisulfide CrossTransportersSynaptic cleftResiduesSame molecule
2008
Mechanism for alternating access in neurotransmitter transporters
Forrest LR, Zhang YW, Jacobs MT, Gesmonde J, Xie L, Honig BH, Rudnick G. Mechanism for alternating access in neurotransmitter transporters. Proceedings Of The National Academy Of Sciences Of The United States Of America 2008, 105: 10338-10343. PMID: 18647834, PMCID: PMC2480614, DOI: 10.1073/pnas.0804659105.Peer-Reviewed Original ResearchConceptsNeurotransmitter transportersMammalian neurotransmitter transportersMammalian serotonin transporterTransmembrane helix 1Bacterial homologueIon-binding sitesTransporter familyExtensive mutagenesisHelix 1Similar repeatsLeuTConformational changesSerotonin transporterRepeatsAlternate conformationConformational differencesExtracellular pathwaysCytoplasmTransportersExtracellular spaceCysteine reagentCrystal structureConformationPathwayAccessibility measurementsInvolvement of serotonin transporter extracellular loop 1 in serotonin binding and transport
Mao Y, Mao Y, Mathewson L, Mao Y, Mathewson L, Gesmonde J, Sato Y, Mao Y, Mathewson L, Gesmonde J, Sato Y, Holy M, Sitte H, Rudnick G. Involvement of serotonin transporter extracellular loop 1 in serotonin binding and transport. Molecular Membrane Biology 2008, 25: 115-127. PMID: 18307099, PMCID: PMC4510095, DOI: 10.1080/09687680701633257.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBiological TransportCell MembraneHeLa CellsHumansIndicators and ReagentsKineticsLigandsMesylatesMolecular Sequence DataMutant ProteinsProtein Structure, TertiaryRatsSequence DeletionSerotoninSerotonin Plasma Membrane Transport ProteinsStructure-Activity Relationship
2000
Permeation and gating residues in serotonin transporter
Chen J, Rudnick G. Permeation and gating residues in serotonin transporter. Proceedings Of The National Academy Of Sciences Of The United States Of America 2000, 97: 1044-1049. PMID: 10655481, PMCID: PMC15515, DOI: 10.1073/pnas.97.3.1044.Peer-Reviewed Original ResearchMeSH KeywordsAllosteric RegulationAmino Acid SubstitutionCarrier ProteinsChloridesCocaineHumansHydrogenIon Channel GatingIon TransportIsoleucineMembrane GlycoproteinsMembrane Transport ProteinsMesylatesMutagenesis, Site-DirectedNerve Tissue ProteinsNorepinephrine Plasma Membrane Transport ProteinsOxidation-ReductionPotassiumProtein BindingProtein ConformationProtein Structure, TertiaryRecombinant Fusion ProteinsReducing AgentsSerotoninSerotonin Plasma Membrane Transport ProteinsSodiumSulfhydryl CompoundsSymportersConceptsSubstrate permeation pathway
1999
The Role of External Loop Regions in Serotonin Transport LOOP SCANNING MUTAGENESIS OF THE SEROTONIN TRANSPORTER EXTERNAL DOMAIN*
Smicun Y, Campbell S, Chen M, Gu H, Rudnick G. The Role of External Loop Regions in Serotonin Transport LOOP SCANNING MUTAGENESIS OF THE SEROTONIN TRANSPORTER EXTERNAL DOMAIN*. Journal Of Biological Chemistry 1999, 274: 36058-36064. PMID: 10593887, DOI: 10.1074/jbc.274.51.36058.Peer-Reviewed Original ResearchConceptsChimeric transportersWild type SERTExternal loop 4High affinity cocaine analogSubsequent conformational changesExternal loop regionsTransmembrane segmentsInitial binding stepScanning mutagenesisWild typeExternal loopLigand bindingSerotonin transporterMutantsConformational changesLoop 4Loop regionConformational flexibilityTransportersCorresponding sequenceBinding stepExternal domainNorepinephrine transporterActivity 5NET substrateMolecular cloning, expression and characterization of a bovine serotonin transporter1The sequence reported in this paper has been deposited in the GenBank data base (accession number AF119122).1
Mortensen O, Kristensen A, Rudnick G, Wiborg O. Molecular cloning, expression and characterization of a bovine serotonin transporter1The sequence reported in this paper has been deposited in the GenBank data base (accession number AF119122).1. Brain Research 1999, 71: 120-126. PMID: 10407194, DOI: 10.1016/s0169-328x(99)00178-3.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsCarrier ProteinsCattleCitalopramCloning, MolecularDesipramineFemaleFluoxetineHeLa CellsHumansImipramineKineticsMembrane GlycoproteinsMembrane Transport ProteinsMolecular Sequence DataNerve Tissue ProteinsN-Methyl-3,4-methylenedioxyamphetamineOrgan SpecificityParoxetinePhylogenyPregnancyRatsRecombinant ProteinsReverse Transcriptase Polymerase Chain ReactionSequence AlignmentSequence Homology, Amino AcidSerotoninSerotonin Plasma Membrane Transport ProteinsTransfectionConceptsSerotonin transporterHuman serotonin transporterExpression of SERTAdrenal glandBrain stemParathyroid glandsPharmacological profileBone marrowThyroid glandSmall intestinePharmacological targetsRT-PCR amplificationDecreased sensitivityExtracellular fluidGlandAmino acid differencesBiogenic aminesNeurotransmitter transportersDependent neurotransmitter transportersImportant antidepressantsAcid differencesDifferent tissuesAntidepressantsParoxetineDesipramine
1998
Critical Amino Acid Residues in Transmembrane Span 7 of the Serotonin Transporter Identified by Random Mutagenesis*
Penado K, Rudnick G, Stephan M. Critical Amino Acid Residues in Transmembrane Span 7 of the Serotonin Transporter Identified by Random Mutagenesis*. Journal Of Biological Chemistry 1998, 273: 28098-28106. PMID: 9774428, DOI: 10.1074/jbc.273.43.28098.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBiological TransportCarrier ProteinsMembrane GlycoproteinsMembrane Transport ProteinsModels, MolecularMolecular Sequence DataMutagenesisNerve Tissue ProteinsProtein ConformationRatsSerotoninSerotonin Plasma Membrane Transport ProteinsStructure-Activity RelationshipConceptsAmino acid residuesRandom mutagenesisAcid residuesTransport activityCritical amino acid residuesRat brain serotonin transporterCritical residuesTransport cycleWild typeNonconservative mutationsStructural predictionsTransporter functionLater stepsMutationsSerotonin transporterResiduesMutagenesisHydrophobic substitutionsTyr-385TransportersMutantsActivitySubstitutionNearby positions
1997
The Third Transmembrane Domain of the Serotonin Transporter Contains Residues Associated with Substrate and Cocaine Binding*
Chen J, Sachpatzidis A, Rudnick G. The Third Transmembrane Domain of the Serotonin Transporter Contains Residues Associated with Substrate and Cocaine Binding*. Journal Of Biological Chemistry 1997, 272: 28321-28327. PMID: 9353288, DOI: 10.1074/jbc.272.45.28321.Peer-Reviewed Original ResearchAsparagineBinding SitesCarrier ProteinsCell LineCell MembraneCocaineCysteineEthyl MethanesulfonateHumansIndicators and ReagentsIsoleucineLigandsMembrane GlycoproteinsMembrane Transport ProteinsMesylatesMutagenesis, Site-DirectedNerve Tissue ProteinsProtein Structure, SecondarySerotoninSerotonin Plasma Membrane Transport ProteinsStructure-Activity RelationshipTyrosinePlacental biogenic amine transporters: cloning and expression
Padbury J, Tseng Y, McGonnigal B, Penado K, Stephan M, Rudnick G. Placental biogenic amine transporters: cloning and expression. Brain Research 1997, 45: 163-168. PMID: 9105686, DOI: 10.1016/s0169-328x(96)00309-9.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBase SequenceBrainCarrier ProteinsCell MembraneCloning, MolecularFemaleHumansMembrane GlycoproteinsMembrane Transport ProteinsMolecular Sequence DataNerve Tissue ProteinsNeuronsPlacentaPregnancyProtein BiosynthesisRecombinant ProteinsSequence Homology, Amino AcidSequence Homology, Nucleic AcidSerotoninSerotonin Plasma Membrane Transport ProteinsSheepAn Extracellular Loop Region of the Serotonin Transporter May Be Involved in the Translocation Mechanism †
Stephan M, Chen M, Penado K, Rudnick G. An Extracellular Loop Region of the Serotonin Transporter May Be Involved in the Translocation Mechanism †. Biochemistry 1997, 36: 1322-1328. PMID: 9063880, DOI: 10.1021/bi962150l.Peer-Reviewed Original ResearchConceptsLarge extracellular loopChimeric transportersWild typeWild type SERTExtracellular loopCocaine analog 2beta-carbomethoxy-3betaCell surface biotinylationWild-type levelsSubstrate translocationExtracellular loop regionSurface biotinylationTranslocation mechanismSerotonin transporterHomologous familyType levelsConformational changesLoop regionRestriction sitesTransportersPoor expressionSubstituted regionsSynaptic cleftDrug bindingSame specificityHigh affinityExternal Cysteine Residues in the Serotonin Transporter †
Chen J, Liu-Chen S, Rudnick G. External Cysteine Residues in the Serotonin Transporter †. Biochemistry 1997, 36: 1479-1486. PMID: 9063896, DOI: 10.1021/bi962256g.Peer-Reviewed Original ResearchConceptsTransport activityMTS reagentsCysteine residuesWild typeWild-type transporterSecond external loopTransient expression systemSurface expressionRat serotonin transporterExternal cysteine residuesHydropathy analysisMutant transportersType transporterDouble mutantExpression systemMethanethiosulfonate reagentsLigand bindingSerotonin transporterMutantsExtracellular loopHeLa cellsDisulfide bondsPartial activityTransportersSerine
1996
Cell-specific Sorting of Biogenic Amine Transporters Expressed in Epithelial Cells*
Gu H, Ahn J, Caplan M, Blakely R, Levey A, Rudnick G. Cell-specific Sorting of Biogenic Amine Transporters Expressed in Epithelial Cells*. Journal Of Biological Chemistry 1996, 271: 18100-18106. PMID: 8663573, DOI: 10.1074/jbc.271.30.18100.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBiogenic AminesBiological TransportCarrier ProteinsCell CompartmentationCell MembraneCell PolarityCells, CulturedDogsDopamineDopamine Plasma Membrane Transport ProteinsEpithelial CellsHumansImmunohistochemistryMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsNorepinephrineNorepinephrine Plasma Membrane Transport ProteinsRatsRecombinant ProteinsSerotoninSerotonin Plasma Membrane Transport ProteinsSymportersConceptsMadin-Darby canine kidneyMDCK cellsLLC-PK1 cellsNeurotransmitter transportersCell surface biotinylationConfocal immunofluorescence microscopyBasolateral membraneCell-specific mechanismsEpithelial cellsBiogenic amine transportersMembrane proteinsSurface biotinylationCDNA encodingHuman DA transporterAmine transportersImmunofluorescence microscopyBiotinylating reagentTransportersPermeable filter supportsApical surfaceImmunocytochemistry resultsBasolateral mediumSurface expressionApical sideDA transporter
1995
Biogenic amine flux mediated by cloned transporters stably expressed in cultured cell lines: amphetamine specificity for inhibition and efflux.
Wall S, Gu H, Rudnick G. Biogenic amine flux mediated by cloned transporters stably expressed in cultured cell lines: amphetamine specificity for inhibition and efflux. Molecular Pharmacology 1995, 47: 544-50. PMID: 7700252.Peer-Reviewed Original ResearchMeSH Keywords1-Methyl-4-phenylpyridiniumAmphetaminesBiogenic MonoaminesBiological TransportCarrier ProteinsCell MembraneCells, CulturedCloning, MolecularCocaineDNA, ComplementaryDopamineDopamine Plasma Membrane Transport ProteinsHumansMazindolMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsNeurotransmitter Uptake InhibitorsNorepinephrineNorepinephrine Plasma Membrane Transport ProteinsSerotoninSerotonin Plasma Membrane Transport ProteinsStimulation, ChemicalSubstrate SpecificitySymportersTransfectionConceptsBiogenic amine transportersCell linesAmine transportersRat serotonin transporterCultured cell linesInhibitor of transportRat dopamine transporterHuman norepinephrine transporterPlasma membraneLLC-PK1 cellsSubstrate effluxSubstrate influxDopamine transporterNorepinephrine transporterAmphetamine derivativesTransportersSerotonin transporterEffluxDistinct patternsP-chloroamphetamineAmine substratesCellsInhibited transportCDNAInhibitors
1994
Ligand binding to the serotonin transporter: equilibria, kinetics, and ion dependence.
Humphreys C, Wall S, Rudnick G. Ligand binding to the serotonin transporter: equilibria, kinetics, and ion dependence. Biochemistry 1994, 33: 9118-25. PMID: 8049215, DOI: 10.1021/bi00197a014.Peer-Reviewed Original Research
1993
Amphetamine derivatives interact with both plasma membrane and secretory vesicle biogenic amine transporters.
Schuldiner S, Steiner-Mordoch S, Yelin R, Wall S, Rudnick G. Amphetamine derivatives interact with both plasma membrane and secretory vesicle biogenic amine transporters. Molecular Pharmacology 1993, 44: 1227-31. PMID: 7903417.Peer-Reviewed Original ResearchMeSH Keywords3,4-MethylenedioxyamphetamineAnimalsBlood PlateletsCarrier ProteinsCattleCell MembraneChromaffin GranulesFenfluramineHumansIn Vitro TechniquesMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsN-Methyl-3,4-methylenedioxyamphetamineP-ChloroamphetamineReserpineSerotoninSerotonin Plasma Membrane Transport ProteinsConceptsVesicular amine transporterVesicular amine transportAmine transportersAmine transportSerotonin transportChromaffin granule membrane vesiclesPlasma membrane transportersBiogenic amine transportersPlasma membraneMembrane transportersMembrane vesiclesSubstrate siteVesicular transportersTransportersDelta pH