Crystal Structures of the Nuclear Receptor, Liver Receptor Homolog 1, Bound to Synthetic Agonists.
Liver receptor homolog 1 (NR5A2, LRH-1) is an orphan nuclear hormone receptor that regulates diverse biological processes, including metabolism, proliferation, and the resolution of endoplasmic reticulum stress. Although preclinical and cellular studies demonstrate that LRH-1 has great potential as a therapeutic target for metabolic diseases and cancer, development of LRH-1 modulators has been difficult. Recently, systematic modifications to one of the few known chemical scaffolds capable of activating LRH-1 failed to improve efficacy substantially. Moreover, mechanisms through which LRH-1 is activated by synthetic ligands are entirely unknown. Here, we use x-ray crystallography and other structural methods to explore conformational changes and receptor-ligand interactions associated with LRH-1 activation by a set of related agonists. Unlike phospholipid LRH-1 ligands, these agonists bind deep in the pocket and do not interact with residues near the mouth nor do they expand the pocket like phospholipids. Unexpectedly, two closely related agonists with similar efficacies (GSK8470 and RJW100) exhibit completely different binding modes. The dramatic repositioning is influenced by a differential ability to establish stable face-to-face π-π-stacking with the LRH-1 residue His-390, as well as by a novel polar interaction mediated by the RJW100 hydroxyl group. The differing binding modes result in distinct mechanisms of action for the two agonists. Finally, we identify a network of conserved water molecules near the ligand-binding site that are important for activation by both agonists. This work reveals a previously unappreciated complexity associated with LRH-1 agonist development and offers insights into rational design strategies.
From the Department of Biochemistry, Emory University School of Medicine, and.
Suzanne G
SG
Mays
From the Department of Biochemistry, Emory University School of Medicine, and.
C Denise
CD
Okafor
the School of Chemistry, University of Southampton, Southampton, Hants SO17 1BJ, United Kingdom, and.
Richard J
RJ
Whitby
the Scripps Research Institute, Jupiter, Florida 33458.
Devrishi
D
Goswami
the School of Chemistry, University of Southampton, Southampton, Hants SO17 1BJ, United Kingdom, and.
Józef
J
Stec
the Department of Chemistry, Emory University, Atlanta, Georgia 30322.
Autumn R
AR
Flynn
the Department of Chemistry, Emory University, Atlanta, Georgia 30322.
Michael C
MC
Dugan
the Department of Chemistry, Emory University, Atlanta, Georgia 30322.
Nathan T
NT
Jui
the Scripps Research Institute, Jupiter, Florida 33458.
Patrick R
PR
Griffin
From the Department of Biochemistry, Emory University School of Medicine, and eortlun@emory.edu.
Eric A
EA
Ortlund
D000814:Aniline Compounds / Q000737:chemistry*; D001643:Bridged Bicyclo Compounds / Q000737:chemistry*; D018360:Crystallography, X-Ray; D006801:Humans; D000072417:Protein Domains; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists* / Q000737:chemistry*
D000814:Aniline Compounds; D001643:Bridged Bicyclo Compounds; C516216:GSK 8470; C082631:NR5A2 protein, human; D018160:Receptors, Cytoplasmic and Nuclear
allosteric regulation; diabetes; lipids; molecular dynamics; nuclear receptor; phosphatidylcholine
J Struct Biol. 2015 Dec;192(3):342-8
26416531
J Mol Graph Model. 2006 Oct;25(2):247-60
16458552
J Am Soc Mass Spectrom. 2013 Oct;24(10):1584-92
23884631
J Clin Invest. 2012 Aug;122(8):2817-26
22772466
Mol Cell Endocrinol. 2004 Feb 27;215(1-2):39-44
15026173
Nucleic Acids Res. 2003 Jul 1;31(13):3352-5
12824325
Protein Eng. 2002 Aug;15(8):697-705
12364585
Cell Metab. 2014 Oct 7;20(4):603-13
25176150
J Biol Chem. 2016 Jan 15;291(3):1411-26
26553876
Mol Cell. 2000 Sep;6(3):507-15
11030331
Nat Struct Mol Biol. 2005 Apr;12(4):357-63
15723037
Endocr Relat Cancer. 2010 Oct 29;17(4):965-75
20817789
Nucleic Acids Res. 2013 Dec;41(22):10228-40
24049078
EMBO Rep. 2002 Dec;3(12):1181-7
12446566
Breast Cancer Res Treat. 2011 Jun;127(2):385-96
20607599
Cancer Res. 2014 Apr 1;74(7):2015-25
24520076
Nat Struct Mol Biol. 2012 Apr 15;19(5):532-7, S1-2
22504882
Stem Cells. 2010 Oct;28(10):1794-804
20734354
J Chem Theory Comput. 2013 Jul 9;9(7):3084-95
26583988
Nat Rev Drug Discov. 2015 Feb;14 (2):95-110
25614222
J Med Chem. 2006 Nov 16;49(23 ):6652-5
17154495
Cancer Lett. 2014 Apr 1;345(1):85-90
24333731
Proc Natl Acad Sci U S A. 2015 Feb 24;112(8):2467-72
25675535
Methods Enzymol. 1997;276:307-26
27754618
Nature. 2011 May 25;474(7352):506-10
21614002
J Med Chem. 2011 Apr 14;54(7):2266-81
21391689
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
20124702
Cell. 2005 Feb 11;120(3):343-55
15707893
Mol Cell Biol. 2005 May;25(9):3492-505
15831456
Biophys J. 2007 Jun 1;92(11):3817-29
17351000
Biophys J. 2007 Sep 1;93(5):1484-92
17483185
J Biol Chem. 2013 Jul 5;288(27):19830-44
23667258
Mol Cell Biol. 2006 Sep;26(18):6799-807
16943422
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
J Am Soc Mass Spectrom. 2012 Sep;23(9):1512-21
22692830
Structure. 2003 Jul;11(7):741-6
12842037
Elife. 2014 Apr 15;3:e01694
24737860
J Am Chem Soc. 2007 Mar 7;129(9):2577-87
17288418
J Chem Inf Model. 2011 Oct 24;51(10):2778-86
21919503
Biochem Biophys Res Commun. 2014 Apr 4;446(2):614-9
24632207