Conformational Dynamics of Human FXR-LBD Ligand Interactions Studied by Hydrogen/deuterium Exchange Mass Spectrometry: Insights into the Antagonism of the Hypolipidemic Agent Z-guggulsterone

Overview

Journal Biochim Biophys Acta

Specialties Biochemistry
Biophysics

Date 2014 Jun 24

PMID 24953769

Citations 10

Authors

Liping Yang

David Broderick

Yuan Jiang

Victor Hsu

Claudia S Maier

Affiliations

Soon will be listed here.

Abstract

Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily of transcription factors that plays a key role in the regulation of bile acids, lipid and glucose metabolisms. The regulative function of FXR is governed by conformational changes of the ligand binding domain (LBD) upon ligand binding. Although FXR is a highly researched potential therapeutic target, only a limited number of FXR-agonist complexes have been successfully crystallized and subsequently yielded high resolution structures. There is currently no structural information of any FXR-antagonist complexes publically available. We therefore explored the use of amide hydrogen/deuterium exchange (HDX) coupled with mass spectrometry for characterizing conformational changes in the FXR-LBD upon ligand binding. Ligand-specific deuterium incorporation profiles were obtained for three FXR ligand chemotypes: GW4064, a synthetic non-steroidal high affinity agonist; the bile acid chenodeoxycholic acid (CDCA), the endogenous low affinity agonist of FXR; and Z-guggulsterone (GG), an in vitro antagonist of the steroid chemotype. A comparison of the HDX profiles of their ligand-bound FXR-LBD complexes revealed a unique mode of interaction for GG. The conformational features of the FXR-LBD-antagonist interaction are discussed.

Citing Articles

On the Cholesterol Raising Effect of Coffee Diterpenes Cafestol and 16--Methylcafestol: Interaction with Farnesoid X Receptor.

Guercia E, Berti F, De Zorzi R, Navarini L, Geremia S, Medagli B Int J Mol Sci. 2024; 25(11).

PMID: 38892285 PMC: 11173301. DOI: 10.3390/ijms25116096.

Exploring the Promising Role of Guggulipid in Rheumatoid Arthritis Management: An In-depth Analysis.

Sahu A, Rathee S, Jain S, Patil U Curr Rheumatol Rev. 2024; 20(5):469-487.

PMID: 38284718 DOI: 10.2174/0115733971280984240101115203.

Conformational restriction shapes the inhibition of a multidrug efflux adaptor protein.

Russell Lewis B, Uddin M, Moniruzzaman M, Kuo K, Higgins A, Shah L Nat Commun. 2023; 14(1):3900.

PMID: 37463890 PMC: 10354078. DOI: 10.1038/s41467-023-39615-x.

Farnesoid X receptor (FXR): Structures and ligands.

Jiang L, Zhang H, Xiao D, Wei H, Chen Y Comput Struct Biotechnol J. 2021; 19:2148-2159.

PMID: 33995909 PMC: 8091178. DOI: 10.1016/j.csbj.2021.04.029.

Dissecting the allosteric FXR modulation: a chemical biology approach using guggulsterone as a chemical tool.

Passeri D, Carotti A, Ramos Pittol J, Ciaccioli G, Pellicciari R, van Mil S Medchemcomm. 2019; 10(8):1412-1419.

PMID: 31673308 PMC: 6786047. DOI: 10.1039/c9md00264b.

References

Huang R, Wen J, Blankenship R, Gross M . Hydrogen-deuterium exchange mass spectrometry reveals the interaction of Fenna-Matthews-Olson protein and chlorosome CsmA protein. Biochemistry. 2011; 51(1):187-93. PMC: 3254703. DOI: 10.1021/bi201620y. View

Soldani C, Bottone M, Pellicciari C, Scovassi A . Nucleolus disassembly in mitosis and apoptosis: dynamic redistribution of phosphorylated-c-Myc, fibrillarin and Ki-67. Eur J Histochem. 2007; 50(4):273-80. View

Ma K, Saha P, Chan L, Moore D . Farnesoid X receptor is essential for normal glucose homeostasis. J Clin Invest. 2006; 116(4):1102-9. PMC: 1409738. DOI: 10.1172/JCI25604. View

Hamuro Y, Coales S, Morrow J, Molnar K, Tuske S, Southern M . Hydrogen/deuterium-exchange (H/D-Ex) of PPARgamma LBD in the presence of various modulators. Protein Sci. 2006; 15(8):1883-92. PMC: 2242592. DOI: 10.1110/ps.062103006. View

Dai S, Burris T, Dodge J, Montrose-Rafizadeh C, Wang Y, Pascal B . Unique ligand binding patterns between estrogen receptor alpha and beta revealed by hydrogen-deuterium exchange. Biochemistry. 2009; 48(40):9668-76. PMC: 2782520. DOI: 10.1021/bi901149t. View

Richter H, Benson G, Bleicher K, Blum D, Chaput E, Clemann N . Optimization of a novel class of benzimidazole-based farnesoid X receptor (FXR) agonists to improve physicochemical and ADME properties. Bioorg Med Chem Lett. 2011; 21(4):1134-40. DOI: 10.1016/j.bmcl.2010.12.123. View

Cui J, Huang L, Zhao A, Lew J, Yu J, Sahoo S . Guggulsterone is a farnesoid X receptor antagonist in coactivator association assays but acts to enhance transcription of bile salt export pump. J Biol Chem. 2003; 278(12):10214-20. DOI: 10.1074/jbc.M209323200. View

Burris T, Montrose C, Houck K, Osborne H, Bocchinfuso W, Yaden B . The hypolipidemic natural product guggulsterone is a promiscuous steroid receptor ligand. Mol Pharmacol. 2004; 67(3):948-54. DOI: 10.1124/mol.104.007054. View

Pellicciari R, Fiorucci S, Camaioni E, Clerici C, Costantino G, Maloney P . 6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), a potent and selective FXR agonist endowed with anticholestatic activity. J Med Chem. 2002; 45(17):3569-72. DOI: 10.1021/jm025529g. View

10.

Sperry J, Huang R, Zhu M, Rempel D, Gross M . Hydrophobic Peptides Affect Binding of Calmodulin and Ca as Explored by H/D Amide Exchange and Mass Spectrometry. Int J Mass Spectrom. 2011; 302(1-3):85-92. PMC: 3134286. DOI: 10.1016/j.ijms.2010.08.013. View

11.

Westin S, Heyman R, Martin R . FXR, a therapeutic target for bile acid and lipid disorders. Mini Rev Med Chem. 2005; 5(8):719-27. DOI: 10.2174/1389557054553802. View

12.

Goodwin B, Jones S, Price R, Watson M, McKee D, Moore L . A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. Mol Cell. 2000; 6(3):517-26. DOI: 10.1016/s1097-2765(00)00051-4. View

13.

Guo G, Santamarina-Fojo S, Akiyama T, Amar M, Paigen B, Brewer Jr B . Effects of FXR in foam-cell formation and atherosclerosis development. Biochim Biophys Acta. 2006; 1761(12):1401-9. PMC: 1751860. DOI: 10.1016/j.bbalip.2006.09.018. View

14.

Han K, Kim J, Kim K, Kim E, Seo J, Yang E . Identification of farnesoid X receptor modulators by a fluorescence polarization-based interaction assay. Anal Biochem. 2009; 398(2):185-90. DOI: 10.1016/j.ab.2009.11.008. View

15.

Weis D, Engen J, Kass I . Semi-automated data processing of hydrogen exchange mass spectra using HX-Express. J Am Soc Mass Spectrom. 2006; 17(12):1700-3. DOI: 10.1016/j.jasms.2006.07.025. View

16.

Englander S, Kallenbach N . Hydrogen exchange and structural dynamics of proteins and nucleic acids. Q Rev Biophys. 1983; 16(4):521-655. DOI: 10.1017/s0033583500005217. View

17.

Hill A . The Combinations of Haemoglobin with Oxygen and with Carbon Monoxide. I. Biochem J. 1913; 7(5):471-80. PMC: 1550542. DOI: 10.1042/bj0070471. View

18.

Zhang Z, Smith D . Determination of amide hydrogen exchange by mass spectrometry: a new tool for protein structure elucidation. Protein Sci. 1993; 2(4):522-31. PMC: 2142359. DOI: 10.1002/pro.5560020404. View

19.

Meyer U, Costantino G, Macchiarulo A, Pellicciari R . Is antagonism of E/Z-guggulsterone at the farnesoid X receptor mediated by a noncanonical binding site? A molecular modeling study. J Med Chem. 2005; 48(22):6948-55. DOI: 10.1021/jm0505056. View

20.

Lundquist J, Harnish D, Kim C, Mehlmann J, Unwalla R, Phipps K . Improvement of physiochemical properties of the tetrahydroazepinoindole series of farnesoid X receptor (FXR) agonists: beneficial modulation of lipids in primates. J Med Chem. 2010; 53(4):1774-87. DOI: 10.1021/jm901650u. View