{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T16:08:04Z","timestamp":1776182884826,"version":"3.50.1"},"reference-count":66,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,18]],"date-time":"2022-12-18T00:00:00Z","timestamp":1671321600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32071233"],"award-info":[{"award-number":["32071233"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2019A1515011569"],"award-info":[{"award-number":["2019A1515011569"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2021A1515012067"],"award-info":[{"award-number":["2021A1515012067"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Guangdong Basic and Applied Basic Research Foundation","award":["32071233"],"award-info":[{"award-number":["32071233"]}]},{"name":"Guangdong Basic and Applied Basic Research Foundation","award":["2019A1515011569"],"award-info":[{"award-number":["2019A1515011569"]}]},{"name":"Guangdong Basic and Applied Basic Research Foundation","award":["2021A1515012067"],"award-info":[{"award-number":["2021A1515012067"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Glycine transporter 1 (GlyT1) is responsible for the reuptake of glycine, which regulates glutamate signaling as a co-agonist with N-methyl-D-aspartic acid (NMDA) receptors in the excitatory synapse and has been proposed to be a potential target in the development of therapies for a broad range of disorders of the central nervous system. Despite significant progress in characterizing structure and transport mechanism of the transporter, the regulation of transport function through oligomerization remains to be understood. In the present work, association of two forms of GlyT1 into dimers and higher order oligomers was detected by coimmunoprecipitation. To investigate functional properties of dimers of a GlyT1 cysteine mutant L288C, we performed oxidative cross-linking of the positioned cysteine residues in extracellular loop 3 (EL3) near the extracellular end of TM6. By analyzing the effect of copper phenanthroline (CuP)-induced dimerization on transport function, cross-linking of L288C was found to inhibit transport activity. In addition, an intramolecular ion pair Lys286-Glu289 was revealed to be critical for stabilizing EL3 in a conformation that modulates CuP-induced dimerization and transport function of the GlyT1 L288C mutant. Furthermore, the influence of transporter conformation on GlyT1 L288C dimerization was investigated. The substrate glycine, in the presence of both Na+ and Cl\u2212, significantly reduced oxidative cross-linking, suggesting a large-scale rotation of the bundle domain during substrate transport impairs interfacial interactions between L288C protomers. The present study provides new insights into structural and functional elements regulating GlyT1 transport activity through its dimerization or oligomerization.<\/jats:p>","DOI":"10.3390\/ijms232416157","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T05:25:46Z","timestamp":1671427546000},"page":"16157","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Cross-Linking and Functional Analyses for Dimerization of a Cysteine Mutant of Glycine Transporter 1"],"prefix":"10.3390","volume":"23","author":[{"given":"Jingru","family":"Wang","sequence":"first","affiliation":[{"name":"School of Life Sciences, Guangzhou University, Guangzhou 510006, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanhe","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Guangzhou University, Guangzhou 510006, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4276-9251","authenticated-orcid":false,"given":"Yuan-Wei","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Guangzhou University, Guangzhou 510006, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"760","DOI":"10.1007\/PL00000899","article-title":"The glycinergic inhibitory synapse","volume":"58","author":"Legendre","year":"2001","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1152\/physrev.00043.2010","article-title":"Fast synaptic inhibition in spinal sensory processing and pain control","volume":"92","author":"Zeilhofer","year":"2012","journal-title":"Physiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/978-3-319-55769-4_4","article-title":"Glycine transporters and its coupling with NMDA receptors","volume":"16","author":"Zafra","year":"2017","journal-title":"Adv. Neurobiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.tibs.2005.04.004","article-title":"Glycine transporters: Essential regulators of neurotransmission","volume":"30","author":"Eulenburg","year":"2005","journal-title":"Trends Biochem. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3336","DOI":"10.1152\/jn.1998.80.6.3336","article-title":"Glycine uptake governs glycine site occupancy at NMDA receptors of excitatory synapses","volume":"80","author":"Berger","year":"1998","journal-title":"J. Neurophysiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.neubiorev.2020.07.025","article-title":"Neurobiology of glycine transporters: From molecules to behavior","volume":"118","author":"Marques","year":"2020","journal-title":"Neurosci. Biobehav. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1517\/13543784.13.5.515","article-title":"The therapeutic potential of glycine transporter-1 inhibitors","volume":"13","author":"Sur","year":"2004","journal-title":"Expert Opin. Investig. Drugs"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.pharmthera.2008.08.004","article-title":"Allosteric modulation of NMDA receptor via elevation of brain glycine and D-serine: The therapeutic potentials for schizophrenia","volume":"120","author":"Yang","year":"2008","journal-title":"Pharmacol. Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.biopsych.2007.09.017","article-title":"Glycine transport inhibitors and the treatment of schizophrenia","volume":"63","author":"Javitt","year":"2008","journal-title":"Biol. Psychiatry"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"866","DOI":"10.1038\/nrd3893","article-title":"Glycine transporters as novel therapeutic targets in schizophrenia, alcohol dependence and pain","volume":"12","author":"Harvey","year":"2013","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1124\/jpet.108.136267","article-title":"Spinal antiallodynia action of glycine transporter inhibitors in neuropathic pain models in mice","volume":"326","author":"Morita","year":"2008","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.pharmthera.2009.03.018","article-title":"Glycine transporter inhibitors as a novel drug discovery strategy for neuropathic pain","volume":"123","author":"Dohi","year":"2009","journal-title":"Pharmacol. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.neulet.2015.06.026","article-title":"Glycine transporter GlyT1, but not GlyT2, is expressed in rat dorsal root ganglion--Possible implications for neuropathic pain","volume":"600","author":"Schlosser","year":"2015","journal-title":"Neurosci. Lett."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1023\/A:1020573325823","article-title":"Bioenergetics of neurotransmitter transport","volume":"30","author":"Rudnick","year":"1998","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"355","DOI":"10.3109\/09687688.2013.842658","article-title":"How do transporters couple solute movements?","volume":"30","author":"Rudnick","year":"2013","journal-title":"Mol. Membr. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1038\/s41467-017-02202-y","article-title":"A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport","volume":"9","author":"Terry","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"E8854","DOI":"10.1073\/pnas.1716870115","article-title":"Structural elements required for coupling ion and substrate transport in the neurotransmitter transporter homolog LeuT","volume":"115","author":"Zhang","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1038\/nature03978","article-title":"Crystal structure of a bacterial homologue of Na+\/Cl\u2212-dependent neurotransmitter transporters","volume":"437","author":"Yamashita","year":"2005","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1038\/nature10737","article-title":"X-ray structures of LeuT in substrate-free outward-open and apo inward-open states","volume":"481","author":"Krishnamurthy","year":"2012","journal-title":"Nature"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"100609","DOI":"10.1016\/j.jbc.2021.100609","article-title":"Crystal structures of LeuT reveal conformational dynamics in the outward-facing states","volume":"296","author":"Fan","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1038\/nature12533","article-title":"X-ray structure of dopamine transporter elucidates antidepressant mechanism","volume":"503","author":"Penmatsa","year":"2013","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1038\/nature14431","article-title":"Neurotransmitter and psychostimulant recognition by the dopamine transporter","volume":"521","author":"Wang","year":"2015","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1038\/s41594-018-0026-8","article-title":"Structural basis for recognition of diverse antidepressants by the human serotonin transporter","volume":"25","author":"Coleman","year":"2018","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1038\/nature17629","article-title":"X-ray structures and mechanism of the human serotonin transporter","volume":"532","author":"Coleman","year":"2016","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1038\/s41586-019-1135-1","article-title":"Serotonin transporter-ibogaine complexes illuminate mechanisms of inhibition and transport","volume":"569","author":"Coleman","year":"2019","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1038\/s41586-021-03274-z","article-title":"Structural insights into the inhibition of glycine reuptake","volume":"591","author":"Shahsavar","year":"2021","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3106","DOI":"10.1073\/pnas.97.7.3106","article-title":"Oligomerization of serotonin transporter and its functional consequences","volume":"97","author":"Kilic","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1046\/j.1471-4159.2003.01793.x","article-title":"Functional consequences of homo- but not hetero-oligomerization between transporters for the biogenic amine neurotransmitters","volume":"85","author":"Kocabas","year":"2003","journal-title":"J. Neurochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1007\/3-540-29784-7_12","article-title":"Oligomerization of neurotransmitter transporters: A ticket from the endoplasmic reticulum to the plasma membrane","volume":"175","author":"Farhan","year":"2006","journal-title":"Handb. Exp. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"14089","DOI":"10.1038\/ncomms14089","article-title":"Direct PIP2 binding mediates stable oligomer formation of the serotonin transporter","volume":"8","author":"Anderluh","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1038\/nature20820","article-title":"The role of interfacial lipids in stabilizing membrane protein oligomers","volume":"541","author":"Gupta","year":"2017","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1111\/jnc.14259","article-title":"Functional properties of dopamine transporter oligomers after copper linking","volume":"144","author":"Zhen","year":"2018","journal-title":"J. Neurochem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"107676","DOI":"10.1016\/j.neuropharm.2019.107676","article-title":"Trimerization of dopamine transporter triggered by AIM-100 binding: Molecular mechanism and effect of mutations","volume":"161","author":"Cheng","year":"2019","journal-title":"Neuropharmacology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5632","DOI":"10.1074\/jbc.RA118.006178","article-title":"Dopamine transporter forms stable dimers in the live cell plasma membrane in a phosphatidylinositol 4,5-bisphosphate-independent manner","volume":"294","author":"Das","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1111\/jnc.15145","article-title":"SLC6 transporter oligomerization","volume":"157","author":"Jayaraman","year":"2021","journal-title":"J. Neurochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"100430","DOI":"10.1016\/j.jbc.2021.100430","article-title":"Direct coupling of oligomerization and oligomerization-driven endocytosis of the dopamine transporter to its conformational mechanics and activity","volume":"296","author":"Sorkina","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4164","DOI":"10.1038\/s41598-021-83374-y","article-title":"Effect of palmitoylation on the dimer formation of the human dopamine transporter","volume":"11","author":"Zeppelin","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"10055","DOI":"10.1073\/pnas.181344298","article-title":"Symmetrical dimer of the human dopamine transporter revealed by cross-linking Cys-306 at the extracellular end of the sixth transmembrane segment","volume":"98","author":"Hastrup","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10978","DOI":"10.1074\/jbc.M800622200","article-title":"Glycine transporter dimers: Evidence for occurrence in the plasma membrane","volume":"283","author":"Bartholomaus","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"12471","DOI":"10.1074\/jbc.M116.763565","article-title":"Allosteric modulation of human dopamine transporter activity under conditions promoting its dimerization","volume":"292","author":"Cheng","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3657","DOI":"10.1021\/acs.jpcb.6b09876","article-title":"Effect of dimerization on the dynamics of neurotransmitter:sodium symporters","volume":"121","author":"Gur","year":"2017","journal-title":"J. Phys. Chem. B"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Jayaraman, K., Morley, A.N., Szollosi, D., Wassenaar, T.A., Sitte, H.H., and Stockner, T. (2018). Dopamine transporter oligomerization involves the scaffold domain, but spares the bundle domain. PLoS Comput. Biol., 14.","DOI":"10.1371\/journal.pcbi.1006229"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"22983","DOI":"10.1074\/jbc.M312484200","article-title":"Zn2+ inhibits glycine transport by glycine transporter subtype 1b","volume":"279","author":"Ju","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4266","DOI":"10.1093\/emboj\/17.15.4266","article-title":"Delineation of an endogenous zinc-binding site in the human dopamine transporter","volume":"17","author":"Norregaard","year":"1998","journal-title":"EMBO J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"36928","DOI":"10.1074\/jbc.274.52.36928","article-title":"Defining proximity relationships in the tertiary structure of the dopamine transporter. Identification of a conserved glutamic acid as a third coordinate in the endogenous Zn2+-binding site","volume":"274","author":"Loland","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/S0014-5793(02)03008-9","article-title":"Zn2+ site engineering at the oligomeric interface of the dopamine transporter","volume":"524","author":"Norregaard","year":"2002","journal-title":"FEBS Lett."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/j.neuropharm.2008.06.016","article-title":"Neurochemical and behavioral profiling of the selective GlyT1 inhibitors ALX5407 and LY2365109 indicate a preferential action in caudal vs. cortical brain areas","volume":"55","author":"Perry","year":"2008","journal-title":"Neuropharmacology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1016\/j.neuropharm.2015.08.031","article-title":"Glycine transporter 1 is a target for the treatment of epilepsy","volume":"99","author":"Shen","year":"2015","journal-title":"Neuropharmacology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1001\/jamapsychiatry.2014.163","article-title":"Effect of bitopertin, a glycine reuptake inhibitor, on negative symptoms of schizophrenia: A randomized, double-blind, proof-of-concept study","volume":"71","author":"Umbricht","year":"2014","journal-title":"JAMA Psychiatry"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"477","DOI":"10.2533\/chimia.2018.477","article-title":"Glycine Transporter type I (GlyT1) inhibitor, bitopertin: A journey from lab to patient","volume":"72","author":"Pinard","year":"2018","journal-title":"Chimia"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1448","DOI":"10.1073\/pnas.98.4.1448","article-title":"Surface-localized glycine transporters 1 and 2 function as monomeric proteins in Xenopus oocytes","volume":"98","author":"Horiuchi","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e2017431118","DOI":"10.1073\/pnas.2017431118","article-title":"Chloride-dependent conformational changes in the GlyT1 glycine transporter","volume":"118","author":"Zhang","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Latka, K., and Bajda, M. (2022). Analysis of binding determinants for different classes of competitive and noncompetitive inhibitors of glycine transporters. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23148050"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"45045","DOI":"10.1074\/jbc.C300349200","article-title":"The human dopamine transporter forms a tetramer in the plasma membrane: Cross-linking of a cysteine in the fourth transmembrane segment is sensitive to cocaine analogs","volume":"278","author":"Hastrup","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2991","DOI":"10.1038\/s41596-018-0075-9","article-title":"Lentiviral transduction of mammalian cells for fast, scalable and high-level production of soluble and membrane proteins","volume":"13","author":"Elegheert","year":"2018","journal-title":"Nat. Protoc."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"28400","DOI":"10.1016\/S0021-9258(18)46941-4","article-title":"Carboxyl terminus of the glycine transporter GLYT1 is necessary for correct processing of the protein","volume":"269","author":"Olivares","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"30807","DOI":"10.1074\/jbc.M504087200","article-title":"Cysteine-scanning mutagenesis of serotonin transporter intracellular loop 2 suggests an alpha-helical conformation","volume":"280","author":"Zhang","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"22926","DOI":"10.1074\/jbc.M312194200","article-title":"Analysis of transmembrane domain 2 of rat serotonin transporter by cysteine scanning mutagenesis","volume":"279","author":"Sato","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"21012","DOI":"10.1074\/jbc.M311972200","article-title":"The role of N-glycosylation in function and surface trafficking of the human dopamine transporter","volume":"279","author":"Li","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1111\/j.1471-4159.2007.05195.x","article-title":"Substrates dissociate dopamine transporter oligomers","volume":"105","author":"Chen","year":"2008","journal-title":"J. Neurochem."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"5.6.1","DOI":"10.1002\/cpbi.3","article-title":"Comparative protein structure modeling using MODELLER","volume":"54","author":"Webb","year":"2016","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2159","DOI":"10.1002\/prot.24403","article-title":"How good is automated protein docking?","volume":"81","author":"Kozakov","year":"2013","journal-title":"Proteins"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"3342","DOI":"10.1093\/bioinformatics\/btw306","article-title":"Accounting for pairwise distance restraints in FFT-based protein-protein docking","volume":"32","author":"Xia","year":"2016","journal-title":"Bioinformatics"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1038\/nprot.2016.169","article-title":"The ClusPro web server for protein-protein docking","volume":"12","author":"Kozakov","year":"2017","journal-title":"Nat. Protoc."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1002\/prot.25219","article-title":"New additions to the ClusPro server motivated by CAPRI","volume":"85","author":"Vajda","year":"2017","journal-title":"Proteins"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1016\/j.str.2020.06.006","article-title":"Performance and its limits in rigid body protein-protein docking","volume":"28","author":"Desta","year":"2020","journal-title":"Structure"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16157\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:43:39Z","timestamp":1760147019000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16157"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,18]]},"references-count":66,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232416157"],"URL":"https:\/\/doi.org\/10.3390\/ijms232416157","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,18]]}}}