Multi-Component Mechanism of H2 Relaxin Binding to RXFP1 through NanoBRET Kinetic Analysis.
作者： Bradley L. Hoare , Shoni Bruell , Ashish Sethi , Paul R. Gooley , Michael J. Lew
DOI: 10.1016/J.ISCI.2018.12.004
关键词:
摘要: The peptide hormone H2 relaxin has demonstrated promise as a therapeutic, but mimetic development been hindered by the poorly understood receptor RXFP1 activation mechanism. is hypothesized to bind two distinct ECD sites, which reorientates N-terminal LDLa module activate transmembrane domain. Here we provide evidence for this model in live cells measuring bioluminescence resonance energy transfer (BRET) between nanoluciferase-tagged constructs and fluorescently labeled (NanoBRET). Additionally, validate these results using related RXFP2 chimeras with an inserted RXFP1-binding domain utilizing NanoBRET nuclear magnetic studies on recombinant proteins. We therefore multi-component molecular mechanism of binding full-length cells. Also, show utility real-time kinetics reveal subtle complexities, may be overlooked traditional equilibrium assays.
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sci-hub.se 参考文章(45)
Leigh A Stoddart, Elizabeth K M Johnstone, Amanda J Wheal, Joëlle Goulding, Matthew B Robers, Thomas Machleidt, Keith V Wood, Stephen J Hill, Kevin D G Pfleger, Application of BRET to monitor ligand binding to GPCRs Nature Methods. ,vol. 12, pp. 661- 663 ,(2015) , 10.1038/NMETH.3398
Zaohua Huang, Courtney Myhr, Ross A. D. Bathgate, Brian A. Ho, Amaya Bueno, Xin Hu, Jingbo Xiao, Noel Southall, Elena Barnaeva, Irina U. Agoulnik, Juan J. Marugan, Marc Ferrer, Alexander I. Agoulnik, Activation of relaxin family receptor 1 from different mammalian species by relaxin peptide and small-molecule agonist ML290 Frontiers in Endocrinology. ,vol. 6, pp. 128- 128 ,(2015) , 10.3389/FENDO.2015.00128
David C Swinney, Paul Beavis, Kai-Ting Chuang, Yue Zheng, Ina Lee, Peter Gee, Jerome Deval, David M Rotstein, Marianna Dioszegi, Palani Ravendran, Jun Zhang, Surya Sankuratri, Rama Kondru, Georges Vauquelin, A study of the molecular mechanism of binding kinetics and long residence times of human CCR5 receptor small molecule allosteric ligands. British Journal of Pharmacology. ,vol. 171, pp. 3364- 3375 ,(2014) , 10.1111/BPH.12683
Erika E. Büllesbach, Christian Schwabe, The trap-like relaxin-binding site of the leucine-rich G-protein-coupled receptor 7. Journal of Biological Chemistry. ,vol. 280, pp. 14051- 14056 ,(2005) , 10.1074/JBC.M500030200
Mary P. Hall, James Unch, Brock F. Binkowski, Michael P. Valley, Braeden L. Butler, Monika G. Wood, Paul Otto, Kristopher Zimmerman, Gediminas Vidugiris, Thomas Machleidt, Matthew B. Robers, Hélène A. Benink, Christopher T. Eggers, Michael R. Slater, Poncho L. Meisenheimer, Dieter H. Klaubert, Frank Fan, Lance P. Encell, Keith V. Wood, Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chemical Biology. ,vol. 7, pp. 1848- 1857 ,(2012) , 10.1021/CB3002478
Daniel J. Scott, Geoffrey W. Tregear, Ross A. D. Bathgate, Modeling the primary hormone-binding site of RXFP1 and RXFP2. Annals of the New York Academy of Sciences. ,vol. 1160, pp. 74- 77 ,(2009) , 10.1111/J.1749-6632.2009.03950.X
Daniel J. Scott, K. Johan Rosengren, Ross A. D. Bathgate, The different ligand-binding modes of relaxin family peptide receptors RXFP1 and RXFP2. Molecular Endocrinology. ,vol. 26, pp. 1896- 1906 ,(2012) , 10.1210/ME.2012-1188
Emma J. Hopkins, Sharon Layfield, Tania Ferraro, Ross A.D. Bathgate, Paul R. Gooley, The NMR solution structure of the relaxin (RXFP1) receptor lipoprotein receptor class A module and identification of key residues in the N-terminal region of the module that mediate receptor activation Journal of Biological Chemistry. ,vol. 282, pp. 4172- 4184 ,(2006) , 10.1074/JBC.M609526200
Peter J. Tummino, Robert A. Copeland, Residence time of receptor-ligand complexes and its effect on biological function. Biochemistry. ,vol. 47, pp. 5481- 5492 ,(2008) , 10.1021/BI8002023
Fazel Shabanpoor, Ross A.D. Bathgate, Alessia Belgi, Linda J. Chan, Vinojini B. Nair, John D. Wade, Mohammed Akhter Hossain, Site-specific conjugation of a lanthanide chelator and its effects on the chemical synthesis and receptor binding affinity of human relaxin-2 hormone. Biochemical and Biophysical Research Communications. ,vol. 420, pp. 253- 256 ,(2012) , 10.1016/J.BBRC.2012.02.141