Hydrogen/deuterium exchange reveals distinct agonist/partial agonist receptor dynamics within vitamin D receptor/retinoid X receptor heterodimer.
Regulation of nuclear receptor (NR) activity is driven by alterations in the conformational dynamics of the receptor upon ligand binding. Previously, we demonstrated that hydrogen/deuterium exchange (HDX) can be applied to determine novel mechanism of action of PPARγ ligands and in predicting tissue specificity of selective estrogen receptor modulators. Here, we applied HDX to probe the conformational dynamics of the ligand binding domain (LBD) of the vitamin D receptor (VDR) upon binding its natural ligand 1α,25-dihydroxyvitamin D3 (1,25D3), and two analogs, alfacalcidol and ED-71. Comparison of HDX profiles from ligands in complex with the LBD with full-length receptor bound to its cognate receptor retinoid X receptor (RXR) revealed unique receptor dynamics that could not be inferred from static crystal structures. These results demonstrate that ligands modulate the dynamics of the heterodimer interface as well as provide insight into the role of AF-2 dynamics in the action of VDR partial agonists.
Zhang
Jun
J
Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida, Jupiter, FL 33458, USA.
Chalmers
Michael J
MJ
Stayrook
Keith R
KR
Burris
Lorri L
LL
Garcia-Ordonez
Ruben D
RD
Pascal
Bruce D
BD
Burris
Thomas P
TP
Dodge
Jeffery A
JA
Griffin
Patrick R
PR
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D002117:Calcitriol / Q000819:agonists / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D018360:Crystallography, X-Ray; D003903:Deuterium / Q000737:chemistry / Q000378:metabolism; D041961:Deuterium Exchange Measurement / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D006859:Hydrogen / Q000737:chemistry / Q000378:metabolism; D006887:Hydroxycholecalciferols / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D055503:Protein Multimerization; D018167:Receptors, Calcitriol / Q000819:agonists / Q000737:chemistry* / Q000378:metabolism; D047488:Retinoid X Receptors / Q000819:agonists / Q000737:chemistry* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D014807:Vitamin D / Q000031:analogs & derivatives
D006887:Hydroxycholecalciferols; D018167:Receptors, Calcitriol; D047488:Retinoid X Receptors; D014807:Vitamin D; D006859:Hydrogen; D003903:Deuterium; D008156:Luciferases; D002117:Calcitriol; C547512:eldecalcitol; C008088:alfacalcidol