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 ResearchMeSH KeywordsAnimalsBinding SitesHumansMolecular Dynamics SimulationPotassiumProtein BindingSerotoninSerotonin Plasma Membrane Transport ProteinsSodiumConceptsSerotonin 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 ResearchMeSH KeywordsAnimalsFluoxetineIbogaineMiceMolecular ConformationSelective Serotonin Reuptake InhibitorsSerotoninSerotonin Plasma Membrane Transport ProteinsSmall Molecule Libraries
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 ResearchMeSH KeywordsAnimalsBiological TransportHumansSerotoninSerotonin Plasma Membrane Transport ProteinsSynaptic Transmission
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 ResearchMeSH KeywordsBiological TransportCocaineCyclic GMP-Dependent Protein KinasesHeLa CellsHumansIbogainePhosphorylationProtein Conformation, alpha-HelicalProtein Processing, Post-TranslationalSerotoninSerotonin Plasma Membrane Transport ProteinsSignal TransductionConceptsTransmembrane 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 ResearchMeSH KeywordsAmino Acid SubstitutionBiological Transport, ActiveCyclic GMPCyclic GMP-Dependent Protein Kinase Type IHEK293 CellsHumansMutation, MissenseP38 Mitogen-Activated Protein KinasesPhosphorylationSerotoninSerotonin Plasma Membrane Transport ProteinsConceptsCGMP-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 cellsThe Mechanistic Basis for Noncompetitive Ibogaine Inhibition of Serotonin and Dopamine Transporters*
Bulling S, Schicker K, Zhang YW, Steinkellner T, Stockner T, Gruber CW, Boehm S, Freissmuth M, Rudnick G, Sitte HH, Sandtner W. The Mechanistic Basis for Noncompetitive Ibogaine Inhibition of Serotonin and Dopamine Transporters*. Journal Of Biological Chemistry 2012, 287: 18524-18534. PMID: 22451652, PMCID: PMC3365767, DOI: 10.1074/jbc.m112.343681.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBinding SitesCell LineDopamine Plasma Membrane Transport ProteinsExcitatory Amino Acid AntagonistsHumansIbogaineKineticsPatch-Clamp TechniquesRadioligand AssaySerotonin Plasma Membrane Transport ProteinsXenopus laevis
2010
Reconstructing a Chloride-binding Site in a Bacterial Neurotransmitter Transporter Homologue*
Tavoulari S, Rizwan AN, Forrest LR, Rudnick G. Reconstructing a Chloride-binding Site in a Bacterial Neurotransmitter Transporter Homologue*. Journal Of Biological Chemistry 2010, 286: 2834-2842. PMID: 21115480, PMCID: PMC3024779, DOI: 10.1074/jbc.m110.186064.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SubstitutionBacterial ProteinsBinding SitesBiological TransportChloridesGram-Positive Endospore-Forming RodsHumansMutation, MissenseNeurotransmitter AgentsSerotonin Plasma Membrane Transport ProteinsStructural Homology, ProteinConceptsChloride-binding siteConformational changesAdjacent binding sitesSingle point mutationProkaryotic homologuesSubstrate translocationIon-binding sitesTransporter homologueTransport proteinsNeurotransmitter transportersNeurotransmitter transportPoint mutationsBinding sitesHomologuesProteinMutationsCl(-) bindsDirect evidenceTherapeutic drugsSitesDependent formTranslocationTransportersBindsResidues
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 ResearchMeSH KeywordsBiological TransportCross-Linking ReagentsCyanogen BromideCysteineCytoplasmHeLa CellsHumansInhibitory Concentration 50KineticsLigandsMembrane Transport ProteinsMutationProtein ConformationSerotoninSerotonin Plasma Membrane Transport ProteinsConceptsN-terminal cyanogen bromide fragmentGamma-aminobutyric acid transporterCyanogen bromide fragmentsTransmembrane 1Cysteine residuesMutagenesis strategyAcid transportersConformational mechanismSerotonin transportCysteineCorresponding positionDisulfide CrossTransportersSynaptic cleftResiduesSame molecule
2008
Involvement 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
2003
Serotonin transporter missense mutation associated with a complex neuropsychiatric phenotype
Ozaki N, Goldman D, Kaye W, Plotnicov K, Greenberg B, Lappalainen J, Rudnick G, Murphy D. Serotonin transporter missense mutation associated with a complex neuropsychiatric phenotype. Molecular Psychiatry 2003, 8: 933-936. PMID: 14593431, DOI: 10.1038/sj.mp.4001365.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnorexia NervosaAsperger SyndromeAutistic DisorderCarrier ProteinsFemaleGenotypeHumansMaleMembrane GlycoproteinsMembrane Transport ProteinsMolecular Sequence DataMutation, MissenseNerve Tissue ProteinsObsessive-Compulsive DisorderPedigreePhenotypePhobic DisordersPolymorphism, Single-Stranded ConformationalProtein Structure, TertiarySerotonin Plasma Membrane Transport Proteins
2001
A Conformationally Sensitive Residue on the Cytoplasmic Surface of Serotonin Transporter*
Androutsellis-Theotokis A, Ghassemi F, Rudnick G. A Conformationally Sensitive Residue on the Cytoplasmic Surface of Serotonin Transporter*. Journal Of Biological Chemistry 2001, 276: 45933-45938. PMID: 11592963, DOI: 10.1074/jbc.m107462200.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCarrier ProteinsCysteineCytoplasmEthyl MethanesulfonateLigandsMembrane GlycoproteinsMembrane Transport ProteinsMutagenesis, Site-DirectedNerve Tissue ProteinsProtein BindingProtein ConformationRatsSerotonin Plasma Membrane Transport ProteinsA Lithium-induced Conformational Change in Serotonin Transporter Alters Cocaine Binding, Ion Conductance, and Reactivity of Cys-109*
Ni Y, Chen J, Androutsellis-Theotokis A, Huang C, Moczydlowski E, Rudnick G. A Lithium-induced Conformational Change in Serotonin Transporter Alters Cocaine Binding, Ion Conductance, and Reactivity of Cys-109*. Journal Of Biological Chemistry 2001, 276: 30942-30947. PMID: 11408487, DOI: 10.1074/jbc.m104653200.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCarrier ProteinsCocaineCysteineGlutamatesHeLa CellsHumansLithiumMembrane GlycoproteinsMembrane PotentialsMembrane Transport ProteinsMesylatesNerve Tissue ProteinsProtein ConformationSerotonin Plasma Membrane Transport ProteinsSodiumXenopus
2000
Functional Role of Critical Stripe Residues in Transmembrane Span 7 of the Serotonin Transporter EFFECTS OF Na+, Li+, AND METHANETHIOSULFONATE REAGENTS*
Kamdar G, Penado K, Rudnick G, Stephan M. Functional Role of Critical Stripe Residues in Transmembrane Span 7 of the Serotonin Transporter EFFECTS OF Na+, Li+, AND METHANETHIOSULFONATE REAGENTS*. Journal Of Biological Chemistry 2000, 276: 4038-4045. PMID: 11058600, DOI: 10.1074/jbc.m008483200.Peer-Reviewed Original ResearchMeSH KeywordsCarrier ProteinsCell MembraneLithiumMembrane GlycoproteinsMembrane Transport ProteinsMesylatesNerve Tissue ProteinsSerotonin Plasma Membrane Transport ProteinsSodiumConceptsCys-109Methanethiosulfonate reagentsWater-filled poresNative cysteine residuesCysteine-containing mutantsExtracellular loop 1Close-contact regionThree-dimensional structureCysteine residuesTranslocation mechanismControl mutantsAlpha-helixMTSEA-biotinResidue positionsCysteine substitutionsLoop 1Conformational changesMTS reagentsFunctional roleMutantsIon bindingResiduesTransportersIon dependenceSerotonin transporterOligomerization of serotonin transporter and its functional consequences
Kilic F, Rudnick G. Oligomerization of serotonin transporter and its functional consequences. Proceedings Of The National Academy Of Sciences Of The United States Of America 2000, 97: 3106-3111. PMID: 10716733, PMCID: PMC16200, DOI: 10.1073/pnas.97.7.3106.Peer-Reviewed Original ResearchBiopolymersCarrier ProteinsGenes, mycHeLa CellsHumansMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsPrecipitin TestsSerotonin Plasma Membrane Transport ProteinsPermeation 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 positionsDetermination of External Loop Topology in the Serotonin Transporter by Site-directed Chemical Labeling*
Chen J, Liu-Chen S, Rudnick G. Determination of External Loop Topology in the Serotonin Transporter by Site-directed Chemical Labeling*. Journal Of Biological Chemistry 1998, 273: 12675-12681. PMID: 9575231, DOI: 10.1074/jbc.273.20.12675.Peer-Reviewed Original ResearchBiotinCarrier ProteinsCysteineLysineMembrane GlycoproteinsMembrane Transport ProteinsMutagenesis, Site-DirectedNerve Tissue ProteinsProtein ConformationSerotonin Plasma Membrane Transport ProteinsSuccinimides
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 RelationshipTyrosine