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dc.contributor.authorPolireddy, Kishore
dc.contributor.authorKhan, Mohiuddin Md. Taimur
dc.contributor.authorChavan, Hemantkumar
dc.contributor.authorYoung, Susan
dc.contributor.authorMa, Xiaochao
dc.contributor.authorWaller, Anna
dc.contributor.authorGarcia, Matthew
dc.contributor.authorPerez, Dominique
dc.contributor.authorChavez, Stephanie
dc.contributor.authorStrouse, Jacob J.
dc.contributor.authorHaynes, Mark K.
dc.contributor.authorBologa, Cristian G.
dc.contributor.authorOprea, Tudor I.
dc.contributor.authorTegos, George
dc.contributor.authorSklar, Larry A.
dc.contributor.authorKrishnamurthy, Partha
dc.date.accessioned2013-03-21T20:40:31Z
dc.date.issued2012
dc.identifier.citationPolireddy, Kishore, Mohiuddin Md. Taimur Khan, Hemantkumar Chavan, Susan Young, Xiaochao Ma, Anna Waller, Matthew Garcia, Dominique Perez, Stephanie Chavez, Jacob J. Strouse, Mark K. Haynes, Cristian G. Bologa, Tudor I. Oprea, George P. Tegos, Larry A. Sklar, and Partha Krishnamurthy. 2012. A novel flow cytometric HTS assay reveals functional modulators of atp binding cassette transporter ABCB6. PLoS ONE 7(7): e40005.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:10456184
dc.description.abstractABCB6 is a member of the adenosine triphosphate (ATP)-binding cassette family of transporter proteins that is increasingly recognized as a relevant physiological and therapeutic target. Evaluation of modulators of ABCB6 activity would pave the way toward a more complete understanding of the significance of this transport process in tumor cell growth, proliferation and therapy-related drug resistance. In addition, this effort would improve our understanding of the function of ABCB6 in normal physiology with respect to heme biosynthesis, and cellular adaptation to metabolic demand and stress responses. To search for modulators of ABCB6, we developed a novel cell-based approach that, in combination with flow cytometric high-throughput screening (HTS), can be used to identify functional modulators of ABCB6. Accumulation of protoporphyrin, a fluorescent molecule, in wild-type ABCB6 expressing K562 cells, forms the basis of the HTS assay. Screening the Prestwick Chemical Library employing the HTS assay identified four compounds, benzethonium chloride, verteporfin, tomatine hydrochloride and piperlongumine, that reduced ABCB6 mediated cellular porphyrin levels. Validation of the identified compounds employing the hemin-agarose affinity chromatography and mitochondrial transport assays demonstrated that three out of the four compounds were capable of inhibiting ABCB6 mediated hemin transport into isolated mitochondria. However, only verteporfin and tomatine hydrochloride inhibited ABCB6’s ability to compete with hemin as an ABCB6 substrate. This assay is therefore sensitive, robust, and suitable for automation in a high-throughput environment as demonstrated by our identification of selective functional modulators of ABCB6. Application of this assay to other libraries of synthetic compounds and natural products is expected to identify novel modulators of ABCB6 activity.en_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi:10.1371/journal.pone.0040005en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3393737/pdf/en_US
dash.licenseLAA
dc.subjectBiologyen_US
dc.subjectBiochemistryen_US
dc.subjectProteinsen_US
dc.subjectTransmembrane Transport Proteinsen_US
dc.subjectDrug Discoveryen_US
dc.subjectBiotechnologyen_US
dc.subjectMolecular Cell Biologyen_US
dc.subjectCytometryen_US
dc.subjectFlow Cytometryen_US
dc.subjectMedicineen_US
dc.subjectDrugs and Devicesen_US
dc.subjectDrug Research and Developmenten_US
dc.titleA Novel Flow Cytometric HTS Assay Reveals Functional Modulators of ATP Binding Cassette Transporter ABCB6en_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS ONEen_US
dash.depositing.authorTegos, George
dc.date.available2013-03-21T20:40:31Z
dc.identifier.doi10.1371/journal.pone.0040005*
dash.contributor.affiliatedTegos, George


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