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
Mephedrone induces partial release at human dopamine transporters but full release at human serotonin transporters
Mayer F, Niello M, Bulling S, Zhang Y, Li Y, Kudlacek O, Holy M, Kooti F, Sandtner W, Rudnick G, Schmid D, Sitte H. Mephedrone induces partial release at human dopamine transporters but full release at human serotonin transporters. Neuropharmacology 2023, 240: 109704. PMID: 37703919, DOI: 10.1016/j.neuropharm.2023.109704.Peer-Reviewed Original ResearchSerotonin transporterCarrier-mediated releaseProtein kinase C inhibitor GF109203X.Human embryonic kidney 293Empathogenic effectsAbuse liabilityGreater efficacyElectrophysiological recordingsDopamine transporterHuman DATReverse transportEntactogenic effectsStimulating propertiesHuman serotonin transporterHuman dopamine transporterMephedroneDopamineMolecular mechanismsHDATReleaserTransportersHigh-affinity transporterDATReleasePrevious studiesStructure-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
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
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 measurements
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 ResearchA 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 Research
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 ResearchConceptsCys-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 transporter
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 DataN-Methyl-3,4-methylenedioxyamphetamineNerve Tissue ProteinsOrgan 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
An 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
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
1993
Binding of the cocaine analog 2 beta-carbomethoxy-3 beta-(4-[125I]iodophenyl)tropane to serotonin and dopamine transporters: different ionic requirements for substrate and 2 beta-carbomethoxy-3 beta-(4-[125I]iodophenyl)tropane binding.
Wall S, Innis R, Rudnick G. Binding of the cocaine analog 2 beta-carbomethoxy-3 beta-(4-[125I]iodophenyl)tropane to serotonin and dopamine transporters: different ionic requirements for substrate and 2 beta-carbomethoxy-3 beta-(4-[125I]iodophenyl)tropane binding. Molecular Pharmacology 1993, 43: 264-70. PMID: 8429827.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBinding, CompetitiveBlood PlateletsCarrier ProteinsCocaineCorpus StriatumDopamineDopamine Plasma Membrane Transport ProteinsHydrogen-Ion ConcentrationIn Vitro TechniquesIonsMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsOsmolar ConcentrationProtein BindingRadioligand AssayRatsSerotoninSerotonin Plasma Membrane Transport ProteinsNon-neurotoxic amphetamine derivatives release serotonin through serotonin transporters.
Rudnick G, Wall S. Non-neurotoxic amphetamine derivatives release serotonin through serotonin transporters. Molecular Pharmacology 1993, 43: 271-6. PMID: 8429828.Peer-Reviewed Original ResearchMeSH KeywordsAmphetaminesBlood PlateletsCarrier ProteinsCell MembraneChromaffin GranulesHumansImipramineIn Vitro TechniquesIndansMembrane GlycoproteinsMembrane Transport ProteinsMethamphetamineModels, BiologicalNerve EndingsNerve Tissue ProteinsRadioligand AssaySerotoninSerotonin Plasma Membrane Transport ProteinsConceptsChromaffin granule membrane vesiclesPlasma membrane vesiclesMembrane vesiclesPlatelet plasma membrane vesiclesPlasma membrane transportersSerotonin transportBiogenic amine transportersMembrane transportersAmine transportersTransportersVesiclesDopamine transporterSerotonin transporterModel systemHalf-maximal concentrationNerve terminalsTransmembraneCocaine analogBindingSimilar concentrationsHigh concentrationsCellsTransport
1992
p-Chloroamphetamine induces serotonin release through serotonin transporters.
Rudnick G, Wall S. p-Chloroamphetamine induces serotonin release through serotonin transporters. Biochemistry 1992, 31: 6710-8. PMID: 1322169, DOI: 10.1021/bi00144a010.Peer-Reviewed Original ResearchMeSH KeywordsBinding, CompetitiveBiological TransportBlood PlateletsCarrier ProteinsCell MembraneChloridesHumansHydrogen-Ion ConcentrationImipramineKineticsLithiumLithium ChlorideMembrane GlycoproteinsMembrane Transport ProteinsNerve Tissue ProteinsP-ChloroamphetamineRadioisotope Dilution TechniqueSerotoninSerotonin Plasma Membrane Transport ProteinsTritiumConceptsPlasma membrane vesiclesMembrane vesiclesChromaffin granule membrane vesiclesVesicular amine transporterATP hydrolysisBovine adrenal chromaffin granulesSerotonin transporterAdrenal chromaffin granulesPrevents accumulationAmine transportersPresence of Mg2TransportersVesiclesChromaffin granulesModel systemDelta pHHuman plateletsATPEffluxManner characteristicAccumulationTransmembraneNaCl gradientPCA's abilityP-chloroamphetamineThe molecular mechanism of "ecstasy" [3,4-methylenedioxy-methamphetamine (MDMA)]: serotonin transporters are targets for MDMA-induced serotonin release.
Rudnick G, Wall S. The molecular mechanism of "ecstasy" [3,4-methylenedioxy-methamphetamine (MDMA)]: serotonin transporters are targets for MDMA-induced serotonin release. Proceedings Of The National Academy Of Sciences Of The United States Of America 1992, 89: 1817-1821. PMID: 1347426, PMCID: PMC48544, DOI: 10.1073/pnas.89.5.1817.Peer-Reviewed Original ResearchConceptsPlasma membrane vesiclesMembrane vesiclesAmine transportersVesicular amine transporterBiogenic amine transportersSecretory vesiclesPlasma membraneATP hydrolysisMolecular mechanismsBovine adrenal chromaffin granulesSerotonin transporterAdrenal chromaffin granulesTransportersDirect interactionVesiclesChromaffin granulesHuman plateletsManner characteristicEffluxTransmembraneATPMDMA actionMechanismBindingMembrane
1989
2-Iodoimipramine, a novel ligand for the serotonin transporter.
Humphreys C, Cassel D, Rudnick G. 2-Iodoimipramine, a novel ligand for the serotonin transporter. Molecular Pharmacology 1989, 36: 620-6. PMID: 2811859.Peer-Reviewed Original Research