PDBsum entry 2aax

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Transcription PDB id
Jmol PyMol
Protein chains
261 a.a. *
PDN ×2
Waters ×443
* Residue conservation analysis
PDB id:
Name: Transcription
Title: Mineralocorticoid receptor double mutant with bound cortisone
Structure: Mineralocorticoid receptor. Chain: a, b. Fragment: ligand binding domain. Synonym: mr. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: nr3c2, mcr, mlr. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
1.75Å     R-factor:   0.185     R-free:   0.212
Authors: R.K.Bledsoe,K.P.Madauss,J.A.Holt,C.J.Apolito,M.H.Lambert, K.H.Pearce,T.B.Stanley,E.L.Stewart,R.P.Trump,T.M.Willson, S.P.Williams
Key ref:
R.K.Bledsoe et al. (2005). A ligand-mediated hydrogen bond network required for the activation of the mineralocorticoid receptor. J Biol Chem, 280, 31283-31293. PubMed id: 15967794 DOI: 10.1074/jbc.M504098200
14-Jul-05     Release date:   26-Jul-05    
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Protein chains
Pfam   ArchSchema ?
P08235  (MCR_HUMAN) -  Mineralocorticoid receptor
984 a.a.
261 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     nucleus   1 term 
  Biological process     steroid hormone mediated signaling pathway   2 terms 
  Biochemical function     DNA binding     2 terms  


DOI no: 10.1074/jbc.M504098200 J Biol Chem 280:31283-31293 (2005)
PubMed id: 15967794  
A ligand-mediated hydrogen bond network required for the activation of the mineralocorticoid receptor.
R.K.Bledsoe, K.P.Madauss, J.A.Holt, C.J.Apolito, M.H.Lambert, K.H.Pearce, T.B.Stanley, E.L.Stewart, R.P.Trump, T.M.Willson, S.P.Williams.
Ligand binding is the first step in hormone regulation of mineralocorticoid receptor (MR) activity. Here, we report multiple crystal structures of MR (NR3C2) bound to both agonist and antagonists. These structures combined with mutagenesis studies reveal that maximal receptor activation involves an intricate ligand-mediated hydrogen bond network with Asn770 which serves dual roles: stabilization of the loop preceding the C-terminal activation function-2 helix and direct contact with the hormone ligand. In addition, most activating ligands hydrogen bond to Thr945 on helix 10. Structural characterization of the naturally occurring S810L mutant explains how stabilization of a helix 3/helix 5 interaction can circumvent the requirement for this hydrogen bond network. Taken together, these results explain the potency of MR activation by aldosterone, the weak activation induced by progesterone and the antihypertensive agent spironolactone, and the binding selectivity of cortisol over cortisone.
  Selected figure(s)  
Figure 3.
FIG. 3. Crystal structure of MR LBD bound to aldosterone and progesterone. A, the overall fold of MR is very similar to the other steroid receptors. B, helix 3 (magenta) residues Asn770 and Ser767 form hydrogen bonds (yellow dashed lines) with the loop (green) residue Glu955 preceding the AF-2 (red). Thr945 present on helix 10 (orange) plays a key role in receptor activation by hydrogen bonding to the C-20 carbonyl and C-21 hydroxyl of aldosterone (yellow). C, close-up view of MR-aldosterone hydrogen bond network. The 18-OH is positioned for hydrogen bonding (yellow dashed lines) with the Asn770 carbonyl, whereas the Asn770 amide remains in position for hydrogen bonding to the C-21 OH of aldosterone and Glu955, which lies in the loop preceding the AF-2. Thr945, present on helix 10, forms a pair of hydrogen bonds with the C-20 and C-21 substituents of aldosterone. Cysteine 942 is in position to interact with the 18-OH group to give aldosterone three potential hydrogen bonds to helix 10. D, progesterone makes no hydrogen bonds to Asn770. Multiple orientations of the Thr945 side chain hydroxyl were observed (A and B) in both noncrystallographically related molecules. When the Thr945 side chain hydroxyl is in the B position (green), the distance (black dashed line) to the progesterone C-20 carbonyl makes hydrogen bonding between the ligand and Thr945 unlikely.
Figure 5.
FIG. 5. Close-up views of the MR C808S/S810L mutant bound to progesterone, cortisone, and spironolactone. A, an overlay of progesterone (yellow) in MR C808S (cyan) and progesterone (pink) in MR C808S/S810L (pink) binding pockets indicates that the S810L mutation has no measurable effect on ligand position or orientation. Hydrogen bonding networks (yellow dashed lines) are indicated. Van der Waals interactions are indicated by black dashed lines. Multiple orientations of the Thr945 side chain hydroxyl were observed (A and B) in both noncrystallographically related molecules. B, the C-11 carbonyl of cortisone (blue) does not interact with MR C808S/S810L (orange) residue Asn770 and is 2.9 Å away from the backbone carbonyl of Leu769. C, the lactone keto group of spironolactone (white) is 3.7 Å away from Asn770 in MR C808S/S810L (magenta), suggesting a weakened potential for hydrogen bonding to Asn770. Again, multiple orientations of the Thr945 side chain hydroxyl were observed (A and B) in both noncrystallographically related molecules. Similar to that observed with progesterone, the distance from the hydroxyl of Thr945 to the lactone keto moiety of spironolactone differs depending upon the orientation of this residue. The weak interaction of spironolactone with both Asn770 and Thr945 is probably the basis for the antagonism observed with this steroid.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2005, 280, 31283-31293) copyright 2005.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20692346 J.J.Kerrigan, Q.Xie, R.S.Ames, and Q.Lu (2011).
Production of protein complexes via co-expression.
  Protein Expr Purif, 75, 1.  
20333619 A.S.Veleiro, L.D.Alvarez, S.L.Eduardo, and G.Burton (2010).
Structure of the glucocorticoid receptor, a flexible protein that can adapt to different ligands.
  ChemMedChem, 5, 649-659.  
20723571 L.Jin, and Y.Li (2010).
Structural and functional insights into nuclear receptor signaling.
  Adv Drug Deliv Rev, 62, 1218-1226.  
20148675 P.Huang, V.Chandra, and F.Rastinejad (2010).
Structural overview of the nuclear receptor superfamily: insights into physiology and therapeutics.
  Annu Rev Physiol, 72, 247-272.  
20817759 R.Goyanka, S.Das, H.H.Samuels, and T.Cardozo (2010).
Nuclear receptor engineering based on novel structure activity relationships revealed by farnesyl pyrophosphate.
  Protein Eng Des Sel, 23, 809-815.  
19114086 C.D.Clyne, C.Y.Chang, R.Safi, P.J.Fuller, D.P.McDonnell, and M.J.Young (2009).
Purification and characterization of recombinant human mineralocorticoid receptor.
  Mol Cell Endocrinol, 302, 81-85.  
19845314 H.Fan, J.J.Irwin, B.M.Webb, G.Klebe, B.K.Shoichet, and A.Sali (2009).
Molecular docking screens using comparative models of proteins.
  J Chem Inf Model, 49, 2512-2527.  
18502379 J.Zhang, and D.S.Geller (2008).
Helix 3-helix 5 interactions in steroid hormone receptor function.
  J Steroid Biochem Mol Biol, 109, 279-285.  
18815806 M.Schubert, F.Brunet, M.Paris, S.Bertrand, G.Benoit, and V.Laudet (2008).
Nuclear hormone receptor signaling in amphioxus.
  Dev Genes Evol, 218, 651-665.  
17904370 N.Yoshimoto, Y.Inaba, S.Yamada, M.Makishima, M.Shimizu, and K.Yamamoto (2008).
2-Methylene 19-nor-25-dehydro-1alpha-hydroxyvitamin D3 26,23-lactones: synthesis, biological activities and molecular basis of passive antagonism.
  Bioorg Med Chem, 16, 457-473.  
17164529 A.M.Hassell, G.An, R.K.Bledsoe, J.M.Bynum, H.L.Carter, S.J.Deng, R.T.Gampe, T.E.Grisard, K.P.Madauss, R.T.Nolte, W.J.Rocque, L.Wang, K.L.Weaver, S.P.Williams, G.B.Wisely, R.Xu, and L.M.Shewchuk (2007).
Crystallization of protein-ligand complexes.
  Acta Crystallogr D Biol Crystallogr, 63, 72-79.  
17702911 E.A.Ortlund, J.T.Bridgham, M.R.Redinbo, and J.W.Thornton (2007).
Crystal structure of an ancient protein: evolution by conformational epistasis.
  Science, 317, 1544-1548.
PDB codes: 2q1h 2q1v 2q3y
17013809 K.P.Madauss, E.L.Stewart, and S.P.Williams (2007).
The evolution of progesterone receptor ligands.
  Med Res Rev, 27, 374-400.  
16972228 L.Pujo, J.Fagart, F.Gary, D.T.Papadimitriou, A.Claës, X.Jeunemaître, and M.C.Zennaro (2007).
Mineralocorticoid receptor mutations are the principal cause of renal type 1 pseudohypoaldosteronism.
  Hum Mutat, 28, 33-40.  
17306029 M.E.Baker, C.Chandsawangbhuwana, and N.Ollikainen (2007).
Structural analysis of the evolution of steroid specificity in the mineralocorticoid and glucocorticoid receptors.
  BMC Evol Biol, 7, 24.  
17206656 S.K.Panigrahi, and G.R.Desiraju (2007).
Strong and weak hydrogen bonds in the protein-ligand interface.
  Proteins, 67, 128-141.  
18174920 S.Viengchareun, D.Le Menuet, L.Martinerie, M.Munier, L.Pascual-Le Tallec, and M.Lombès (2007).
The mineralocorticoid receptor: insights into its molecular and (patho)physiological biology.
  Nucl Recept Signal, 5, e012.  
16892386 J.T.Moore, J.L.Collins, and K.H.Pearce (2006).
The nuclear receptor superfamily and drug discovery.
  ChemMedChem, 1, 504-523.  
16413609 O.Skøtt, T.R.Uhrenholt, J.Schjerning, P.B.Hansen, L.E.Rasmussen, and B.L.Jensen (2006).
Rapid actions of aldosterone in vascular health and disease--friend or foe?
  Pharmacol Ther, 111, 495-507.  
16503757 P.Fuller (2006).
The aldosterone receptor--new insights?
  Expert Opin Investig Drugs, 15, 201-203.  
16722436 S.Inaba, and I.Miyamori (2006).
[Hypertension due to mutation of mineralocorticoid receptors]
  Nippon Naika Gakkai Zasshi, 95, 677-682.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.