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dc.contributor.authorHoare, BL
dc.contributor.authorBruell, S
dc.contributor.authorSethi, A
dc.contributor.authorGooley, PR
dc.contributor.authorLew, MJ
dc.contributor.authorHossain, MA
dc.contributor.authorInoue, A
dc.contributor.authorScott, DJ
dc.contributor.authorBathgate, RAD
dc.date.accessioned2020-12-10T01:06:19Z
dc.date.available2020-12-10T01:06:19Z
dc.date.issued2019-01-25
dc.identifierpii: S2589-0042(18)30233-5
dc.identifier.citationHoare, B. L., Bruell, S., Sethi, A., Gooley, P. R., Lew, M. J., Hossain, M. A., Inoue, A., Scott, D. J. & Bathgate, R. A. D. (2019). Multi-Component Mechanism of H2 Relaxin Binding to RXFP1 through NanoBRET Kinetic Analysis. ISCIENCE, 11, pp.93-+. https://doi.org/10.1016/j.isci.2018.12.004.
dc.identifier.issn2589-0042
dc.identifier.urihttp://hdl.handle.net/11343/253658
dc.description.abstractThe peptide hormone H2 relaxin has demonstrated promise as a therapeutic, but mimetic development has been hindered by the poorly understood relaxin receptor RXFP1 activation mechanism. H2 relaxin is hypothesized to bind to two distinct ECD sites, which reorientates the N-terminal LDLa module to activate the transmembrane domain. Here we provide evidence for this model in live cells by measuring bioluminescence resonance energy transfer (BRET) between nanoluciferase-tagged RXFP1 constructs and fluorescently labeled H2 relaxin (NanoBRET). Additionally, we validate these results using the related RXFP2 receptor and chimeras with an inserted RXFP1-binding domain utilizing NanoBRET and nuclear magnetic resonance studies on recombinant proteins. We therefore provide evidence for the multi-component molecular mechanism of H2 relaxin binding to RXFP1 on the full-length receptor in cells. Also, we show the utility of NanoBRET real-time binding kinetics to reveal subtle binding complexities, which may be overlooked in traditional equilibrium binding assays.
dc.languageEnglish
dc.publisherCELL PRESS
dc.titleMulti-Component Mechanism of H2 Relaxin Binding to RXFP1 through NanoBRET Kinetic Analysis
dc.typeJournal Article
dc.identifier.doi10.1016/j.isci.2018.12.004
melbourne.affiliation.departmentFlorey Department of Neuroscience and Mental Health
melbourne.affiliation.departmentBiochemistry and Molecular Biology
melbourne.affiliation.departmentPharmacology and Therapeutics
melbourne.source.titleiScience
melbourne.source.volume11
melbourne.source.pages93-+
dc.rights.licenseCC BY-NC-ND
melbourne.elementsid1364300
melbourne.contributor.authorSethi, Ashish
melbourne.contributor.authorBathgate, Ross
melbourne.contributor.authorScott, Daniel
melbourne.contributor.authorHossain, Mohammed
melbourne.contributor.authorLew, Michael
melbourne.contributor.authorGooley, Paul
melbourne.contributor.authorBruell, Shoni
melbourne.contributor.authorBruell, Shoni
dc.identifier.eissn2589-0042
melbourne.accessrightsOpen Access


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