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 ResearchSLC6 neurotransmitter transporter family in GtoPdb v.2023.1
Bröer S, Rudnick G. SLC6 neurotransmitter transporter family in GtoPdb v.2023.1. IUPHAR/BPS Guide To Pharmacology CITE 2023, 2023 DOI: 10.2218/gtopdb/f144/2023.1.Peer-Reviewed Original ResearchSolute carrier family 6Transporter familyNeurotransmitter transporter familyDependent neurotransmitter transportersAmino acid transportersDependent amino acid transporterNSS transportersTM segmentsPlasma membraneNeurotransmitter transportersAcid transportersFamily 6Neutral amino acidsAmino acidsTransportersStructural motifsGtoPdb v.LeuTAeolicusSubfamiliesFamilyLeuTAaMembersMotifCrystal structure
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 ResearchSLC6 neurotransmitter transporter family (version 2019.4) in the IUPHAR/BPS Guide to Pharmacology Database
Bröer S, Rudnick G. SLC6 neurotransmitter transporter family (version 2019.4) in the IUPHAR/BPS Guide to Pharmacology Database. IUPHAR/BPS Guide To Pharmacology CITE 2019, 2019 DOI: 10.2218/gtopdb/f144/2019.4.Peer-Reviewed Reviews, Practice Guidelines, Standards, and Consensus StatementsSolute carrier family 6Transporter familyNeurotransmitter transporter familyDependent neurotransmitter transportersAmino acid transportersDependent amino acid transporterNSS transportersTM segmentsPlasma membraneNeurotransmitter transportersAcid transportersFamily 6Neutral amino acidsIUPHAR/BPS GuideAmino acidsTransportersStructural motifsLeuTAeolicusSubfamiliesFamilyPharmacology DatabaseLeuTAaMembersMotif
2018
Structural elements required for coupling ion and substrate transport in the neurotransmitter transporter homolog LeuT
Zhang YW, Tavoulari S, Sinning S, Aleksandrova AA, Forrest LR, Rudnick G. Structural elements required for coupling ion and substrate transport in the neurotransmitter transporter homolog LeuT. Proceedings Of The National Academy Of Sciences Of The United States Of America 2018, 115: e8854-e8862. PMID: 30181291, PMCID: PMC6156673, DOI: 10.1073/pnas.1716870115.Peer-Reviewed Original ResearchConceptsTransporter domainConformational changesOpen stateSodium symporter familyIon-substrate couplingTransmembrane ion gradientsSymporter familyNSS transportersSubstrate bindingLeuTIntracellular substratesCysteine accessibilitySubstrate transportAccessibility of substrateTyrosine residuesConformational responseNa2 siteUncoupled movementIon gradientsExtracellular pathwaysMechanistic componentsTransportersProteinTransport of ionsBinding
2017
Substrate-Induced Conformational Change in LeuT
Zhang Y, Forrest L, Rudnick G. Substrate-Induced Conformational Change in LeuT. Biophysical Journal 2017, 112: 129a. DOI: 10.1016/j.bpj.2016.11.717.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
2015
Two Na+ Sites Control Conformational Change in a Neurotransmitter Transporter Homolog*
Tavoulari S, Margheritis E, Nagarajan A, DeWitt DC, Zhang YW, Rosado E, Ravera S, Rhoades E, Forrest LR, Rudnick G. Two Na+ Sites Control Conformational Change in a Neurotransmitter Transporter Homolog*. Journal Of Biological Chemistry 2015, 291: 1456-1471. PMID: 26582198, PMCID: PMC4714228, DOI: 10.1074/jbc.m115.692012.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SubstitutionAmino Acid Transport SystemsAquatic OrganismsBacterial ProteinsBinding SitesCysteineGram-Negative BacteriaLigandsLiposomesModels, MolecularMolecular Dynamics SimulationMutagenesis, Site-DirectedMutationPlasma Membrane Neurotransmitter Transport ProteinsProtein ConformationProtein FoldingProtein StabilityProteolipidsRecombinant ProteinsSodiumConceptsConformational changesTransmembrane helix 1Open conformational stateDependent conformational changesTransporter homologExtracellular gateProkaryotic homologCytoplasmic pathwayHelix 1Interaction networksIntermediary interactionsBiophysical assaysNeurotransmitter transportersSubstrate pathwayNa2 siteConformational statesHelix motionsLeuTDirect interactionDependent closureHomologMutantsDistinct stepsResiduesComputational analysis
2014
The Role of Sodium Sites in LeuT Conformational Changes
Rudnick G, Tavoulari S, Zhang Y, DeWitt D, Nagarajan A, Rosado E, Ravera S, Kreuder A, Forrest L, Rhoades E. The Role of Sodium Sites in LeuT Conformational Changes. Biophysical Journal 2014, 106: 228a-229a. DOI: 10.1016/j.bpj.2013.11.1337.Peer-Reviewed Original Research
2013
How do transporters couple solute movements?
Rudnick G. How do transporters couple solute movements? Molecular Membrane Biology 2013, 30: 355-359. PMID: 24147977, PMCID: PMC4077868, DOI: 10.3109/09687688.2013.842658.Peer-Reviewed Reviews, Practice Guidelines, Standards, and Consensus StatementsIsoforms of the neuronal glutamate transporter gene, SLC1A1/EAAC1, negatively modulate glutamate uptake: relevance to obsessive-compulsive disorder
Porton B, Greenberg B, Askland K, Serra L, Gesmonde J, Rudnick G, Rasmussen S, Kao H. Isoforms of the neuronal glutamate transporter gene, SLC1A1/EAAC1, negatively modulate glutamate uptake: relevance to obsessive-compulsive disorder. Translational Psychiatry 2013, 3: e259-e259. PMID: 23695234, PMCID: PMC3669922, DOI: 10.1038/tp.2013.35.Peer-Reviewed Original ResearchConceptsExcellent candidate geneObsessive-compulsive disorderNeuronal glutamate transporterGenomic levelInternal promoterCandidate genesPartial colocalizationTransporter geneGene expressionGenetic studiesGlutamate uptakeEAAC1 transporterPrimary promoterEAAC1 proteinGlutamate transporter geneGenesPhysiological regulatorIsoformsTranscriptsExon 2Glutamate transportersGlutamatergic gene expressionPromoterPotential clinical utilityNeuronal glutamate transporter gene
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 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 ResearchA Unifying Concept of Serotonin Transporter Associated Currents
Sandtner W, Schicker K, Freissmuth M, Rudnick G, Sitte H. A Unifying Concept of Serotonin Transporter Associated Currents. Biophysical Journal 2012, 102: 714a. DOI: 10.1016/j.bpj.2011.11.3875.Peer-Reviewed Original Research
2011
Cytoplasmic Permeation Pathway of Neurotransmitter Transporters
Rudnick G. Cytoplasmic Permeation Pathway of Neurotransmitter Transporters. Biochemistry 2011, 50: 7462-7475. PMID: 21774491, PMCID: PMC3164596, DOI: 10.1021/bi200926b.Peer-Reviewed Original ResearchConceptsCytoplasmic permeation pathwaysBacterial amino acid transporter LeuTNeurotransmitter transportersPermeation pathwayKingdoms of lifeMammalian serotonin transporterHigh-resolution crystal structuresFour-helix bundleRecent high-resolution crystal structureSubsequent crystal structureStructural repeatsLeuT structureProtein familyCommon structural featuresSolute transportersRelated proteinsLarge structural familyCytoplasmic oneConformational changesSubstrate siteFirst structureBiological membranesTransportersLeuTExtracellular pathways
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 ResearchConceptsChloride-binding siteConformational changesAdjacent binding sitesSingle point mutationProkaryotic homologuesSubstrate translocationIon-binding sitesTransporter homologueTransport proteinsNeurotransmitter transportersNeurotransmitter transportPoint mutationsBinding sitesHomologuesProteinMutationsCl(-) bindsDirect evidenceTherapeutic drugsSitesDependent formTranslocationTransportersBindsResiduesIbogaine Stimulates Ionic Currents Mediated by Serotonin Transporter
Zhang W, Tao Z, Gesmonde J, Howe J, Rudnick G. Ibogaine Stimulates Ionic Currents Mediated by Serotonin Transporter. Biophysical Journal 2010, 98: 505a. DOI: 10.1016/j.bpj.2009.12.2749.Peer-Reviewed Original Research
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