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dc.contributor.authorTavoulari, Sotiria
dc.contributor.authorSchirris, Tom JJ
dc.contributor.authorMavridou, Vasiliki
dc.contributor.authorThangaratnarajah, Chancievan
dc.contributor.authorKing, Martin S
dc.contributor.authorJones, Daniel TD
dc.contributor.authorDing, Shujing
dc.contributor.authorFearnley, Ian M
dc.contributor.authorKunji, Edmund
dc.date.accessioned2022-04-13T01:02:20Z
dc.date.available2022-04-13T01:02:20Z
dc.date.issued2022-06
dc.identifier.issn2212-8778
dc.identifier.other35278701
dc.identifier.otherPMC8968063
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/336045
dc.description.abstractOBJECTIVE: The mitochondrial pyruvate carrier (MPC) has emerged as a promising drug target for metabolic disorders, including non-alcoholic steatohepatitis and diabetes, metabolically dependent cancers and neurodegenerative diseases. A range of structurally diverse small molecule inhibitors have been proposed, but the nature of their interaction with MPC is not understood, and the composition of the functional human MPC is still debated. The goal of this study was to characterise the human MPC protein in vitro, to understand the chemical features that determine binding of structurally diverse inhibitors and to develop novel higher affinity ones. METHODS: We recombinantly expressed and purified human MPC hetero-complexes and studied their composition, transport and inhibitor binding properties by establishing in vitro transport assays, high throughput thermostability shift assays and pharmacophore modeling. RESULTS: We determined that the functional unit of human MPC is a hetero-dimer. We compared all different classes of MPC inhibitors to find that three closely arranged hydrogen bond acceptors followed by an aromatic ring are shared characteristics of all inhibitors and represent the minimal requirement for high potency. We also demonstrated that high affinity binding is not attributed to covalent bond formation with MPC cysteines, as previously proposed. Following the basic pharmacophore properties, we identified 14 new inhibitors of MPC, one outperforming compound UK5099 by tenfold. Two are the commonly prescribed drugs entacapone and nitrofurantoin, suggesting an off-target mechanism associated with their adverse effects. CONCLUSIONS: This work defines the composition of human MPC and the essential MPC inhibitor characteristics. In combination with the functional assays we describe, this new understanding will accelerate the development of clinically relevant MPC modulators.
dc.languageeng
dc.publisherElsevier BV
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.sourcenlmid: 101605730
dc.sourceessn: 2212-8778
dc.subjectMitochondria
dc.subjectInhibition
dc.subjectSmall molecules
dc.subjectMitochondrial Transport
dc.subjectMitochondrial Pyruvate Carrier
dc.titleKey features of inhibitor binding to the human mitochondrial pyruvate carrier hetero-dimer.
dc.typeArticle
dc.date.updated2022-04-13T01:02:11Z
prism.publicationNameMol Metab
prism.volume60
dc.identifier.doi10.17863/CAM.83474
dcterms.dateAccepted2022-03-01
rioxxterms.versionofrecord10.1016/j.molmet.2022.101469
rioxxterms.versionVoR
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidKunji, Edmund [0000-0002-0610-4500]
dc.identifier.eissn2212-8778
pubs.funder-project-idCancer Research UK (21617)
pubs.funder-project-idMedical Research Council (MC_UU_00015/1)


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International