PDBsum entry 1t65

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Hormone/growth factor PDB id
Jmol PyMol
Protein chain
251 a.a. *
Waters ×320
* Residue conservation analysis
PDB id:
Name: Hormone/growth factor
Title: Crystal structure of the androgen receptor ligand binding domain with dht and a peptide derived form its physiological coactivator grip1 nr box 2 bound in a non- helical conformation
Structure: Androgen receptor. Chain: a. Fragment: ar ligand binding domain 669-919. Synonym: dihydrotestosterone receptor. Engineered: yes. Nuclear receptor coactivator 2. Chain: b. Synonym: ncoa-2, transcriptional intermediary factor 2. Engineered: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: ar, nr3c4, dhtr. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: ncoa2, tif2. Expression_system_taxid: 562
Biol. unit: Dimer (from PQS)
1.66Å     R-factor:   0.230     R-free:   0.270
Authors: E.Estebanez-Perpina,J.M.R.Moore,E.Mar,P.Nguyen,E.Delgado- Rodrigues,J.D.Baxter,P.Webb,R.J.Fletterick,R.K.Guy
Key ref:
E.Estébanez-Perpiñá et al. (2005). The molecular mechanisms of coactivator utilization in ligand-dependent transactivation by the androgen receptor. J Biol Chem, 280, 8060-8068. PubMed id: 15563469 DOI: 10.1074/jbc.M407046200
05-May-04     Release date:   25-Jan-05    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P10275  (ANDR_HUMAN) -  Androgen receptor
919 a.a.
251 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 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.M407046200 J Biol Chem 280:8060-8068 (2005)
PubMed id: 15563469  
The molecular mechanisms of coactivator utilization in ligand-dependent transactivation by the androgen receptor.
E.Estébanez-Perpiñá, J.M.Moore, E.Mar, E.Delgado-Rodrigues, P.Nguyen, J.D.Baxter, B.M.Buehrer, P.Webb, R.J.Fletterick, R.K.Guy.
Androgens drive sex differentiation, bone and muscle development, and promote growth of hormone-dependent cancers by binding the nuclear androgen receptor (AR), which recruits coactivators to responsive genes. Most nuclear receptors recruit steroid receptor coactivators (SRCs) to their ligand binding domain (LBD) using a leucine-rich motif (LXXLL). AR is believed to recruit unique coactivators to its LBD using an aromatic-rich motif (FXXLF) while recruiting SRCs to its N-terminal domain (NTD) through an alternate mechanism. Here, we report that the AR-LBD interacts with both FXXLF motifs and a subset of LXXLL motifs and that contacts with these LXXLL motifs are both necessary and sufficient for SRC-mediated AR regulation of transcription. Crystal structures of the activated AR in complex with both recruitment motifs reveal that side chains unique to the AR-LBD rearrange to bind either the bulky FXXLF motifs or the more compact LXXLL motifs and that AR utilizes subsidiary contacts with LXXLL flanking sequences to discriminate between LXXLL motifs.
  Selected figure(s)  
Figure 3.
FIG. 3. Associations of the AR-LBD with coactivator domains determined by x-ray crystallography (A-H). Close-up views of the interaction between ARA70, SRC2-3, SRC2-2, and SRC3-2 peptides with AR-LBD AF2. The nuclear receptor AF-2 transactivation function is ascribed to a surface-exposed hydrophobic cleft comprising residues from helices 3 (H3, dark blue), 5 (H5, pale blue), and 12 (H12, red), as can be clearly seen in the bottom figures (E-H). A-H, the helix backbone of peptides from ARA70 (RETSEKFKLLFQSYN) (left, red), SRC2-3 (KENALLRYLLDKDD) (middle left, yellow), and SRC3-2 (HKKLLQLLT) (middle right, orange) are shown, and the non-helical SRC2-2 peptide backbone (KHKILHRLLQDSS) (right, green) can be seen. AR-LBD is represented by a solid semi-transparent surface (gray)inthe top figures (A-D). The side chains of the motif hydrophobic residues Phe+1/Leu+1, Leu+4, and Phe+5/Leu+5 of the peptides are shown as stick models. Helix 12, with its Glu-897 side chain, stabilizes the N terminus of the ARA70 peptide, but not those of the SRC peptides. On H3, the side chain of Lys-720 is shown capping the C terminus of ARA70 and SRC2-3 peptides (E and F). B, the side chains of the AR-LBD residues contacting the peptides are depicted as stick models. ARA70: The triad compressed by the Phe aromatic side chains and Leu+4(FXXLF) fits tightly into a deep narrow pocket comprised of Val-716 and Val-730, Met-734, Ile-737, and the hydrophobic segment of Glu-893. The Leu side chains of SRC2-3 and SRC3-2 fit loosely into a flat hydrophobic pocket comprising the side chains of three valines, 716, 730, and 901, methionines 734 and 894, glutamines 733 and 738, Asp-731, and Glu-897. The accommodation of the bulkier Phe residues of ARA70 is accompanied by the rearrangements of Met-734, Glu-897, and Lys-720 predominantly (indicated by gray dots on the surface representation of AR). D and H, SRC2-2 does not bind to AR-LBD AF2 in an helical conformation, and, apart from Leu+1, the rest of the peptide cannot be superimposed to the other SRC peptides shown in this report. All the figures were generated with Pymol (42).
Figure 4.
FIG. 4. Role of the binding pocket and charge clamp residues of the AR-LBD AF-2 in interaction with cofactors and potentiation of transcriptional activation by a GAL4-AR-LBD construct. A, removing the charge at Lys-720 or reversing the charge at Glu-897 (the positive and negative ends of the "charge clamp" that stabilizes helix dipole for the NR box) markedly reduces the potentiation of transcriptional activation by GAL AR-LBD by SRC2 in HeLa cells. However, neutralization of the charge at Glu-897 has modest effects on transcriptional activation. B, Western blot demonstrating that all Glu-897 mutants are expressed at similar levels in HeLa cells during the transactivation experiments. C, as expected from the peptide binding data (Fig. 1A), neutralization of charge at Glu-897 has no discernable effect upon the interaction of SRC2 as measured by GST Pull-down. Similarly, there is a modest reduction in binding of AR NTD by E897Q. However, reversal of charge (E897K) strongly reduces binding of both SRC2 and AR NTD.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2005, 280, 8060-8068) copyright 2005.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20690138 D.J.van de Wijngaart, H.J.Dubbink, M.Molier, Vos, G.Jenster, and J.Trapman (2011).
Inhibition of androgen receptor functions by gelsolin FxxFF peptide delivered by transfection, cell-penetrating peptides, and lentiviral infection.
  Prostate, 71, 241-253.  
20011049 A.T.Szafran, S.Hartig, H.Sun, I.P.Uray, M.Szwarc, Y.Shen, S.N.Mediwala, J.Bell, M.J.McPhaul, M.A.Mancini, and M.Marcelli (2009).
Androgen receptor mutations associated with androgen insensitivity syndrome: a high content analysis approach leading to personalized medicine.
  PLoS One, 4, e8179.  
19183053 A.Teichert, L.A.Arnold, S.Otieno, Y.Oda, I.Augustinaite, T.R.Geistlinger, R.W.Kriwacki, R.K.Guy, and D.D.Bikle (2009).
Quantification of the vitamin D receptor-coregulator interaction.
  Biochemistry, 48, 1454-1461.  
  19645433 C.Féau, L.A.Arnold, A.Kosinski, F.Zhu, M.Connelly, and R.K.Guy (2009).
Novel flufenamic acid analogues as inhibitors of androgen receptor mediated transcription.
  ACS Chem Biol, 4, 834-843.  
19126541 J.Fu, J.Jiang, J.Li, S.Wang, G.Shi, Q.Feng, E.White, J.Qin, and J.Wong (2009).
Deleted in breast cancer 1, a novel androgen receptor (AR) coactivator that promotes AR DNA-binding activity.
  J Biol Chem, 284, 6832-6840.  
  19441848 J.R.Gunther, A.A.Parent, and J.A.Katzenellenbogen (2009).
Alternative inhibition of androgen receptor signaling: peptidomimetic pyrimidines as direct androgen receptor/coactivator disruptors.
  ACS Chem Biol, 4, 435-440.  
18037956 G.N.Brooke, M.G.Parker, and C.L.Bevan (2008).
Mechanisms of androgen receptor activation in advanced prostate cancer: differential co-activator recruitment and gene expression.
  Oncogene, 27, 2941-2950.  
18852122 M.C.Hodgson, H.C.Shen, A.N.Hollenberg, and S.P.Balk (2008).
Structural basis for nuclear receptor corepressor recruitment by antagonist-liganded androgen receptor.
  Mol Cancer Ther, 7, 3187-3194.  
18668523 P.Singh, G.Hallur, R.K.Anchoori, O.Bakare, Y.Kageyama, S.R.Khan, and J.T.Isaacs (2008).
Rational design of novel antiandrogens for neutralizing androgen receptor function in hormone refractory prostate cancer.
  Prostate, 68, 1570-1581.  
18593950 S.M.Dehm, L.J.Schmidt, H.V.Heemers, R.L.Vessella, and D.J.Tindall (2008).
Splicing of a novel androgen receptor exon generates a constitutively active androgen receptor that mediates prostate cancer therapy resistance.
  Cancer Res, 68, 5469-5477.  
17911242 E.Estébanez-Perpiñá, L.A.Arnold, A.A.Arnold, P.Nguyen, E.D.Rodrigues, E.Mar, R.Bateman, P.Pallai, K.M.Shokat, J.D.Baxter, R.K.Guy, P.Webb, and R.J.Fletterick (2007).
A surface on the androgen receptor that allosterically regulates coactivator binding.
  Proc Natl Acad Sci U S A, 104, 16074-16079.
PDB codes: 2pio 2pip 2piq 2pir 2pit 2piu 2piv 2piw 2pix 2pkl 2qpy
17300979 V.Nahoum, and W.Bourguet (2007).
Androgen and estrogen receptors: potential of crystallography in the fight against cancer.
  Int J Biochem Cell Biol, 39, 1280-1287.  
17606915 W.H.Bisson, A.V.Cheltsov, N.Bruey-Sedano, B.Lin, J.Chen, N.Goldberger, L.T.May, A.Christopoulos, J.T.Dalton, P.M.Sexton, X.K.Zhang, and R.Abagyan (2007).
Discovery of antiandrogen activity of nonsteroidal scaffolds of marketed drugs.
  Proc Natl Acad Sci U S A, 104, 11927-11932.  
  17205105 B.Comuzzi, and M.D.Sadar (2006).
Proteomic analyses to identify novel therapeutic targets for the treatment of advanced prostate cancer.
  Cellscience, 3, 61-81.  
16641486 K.Pereira de Jésus-Tran, P.L.Côté, L.Cantin, J.Blanchet, F.Labrie, and R.Breton (2006).
Comparison of crystal structures of human androgen receptor ligand-binding domain complexed with various agonists reveals molecular determinants responsible for binding affinity.
  Protein Sci, 15, 987-999.
PDB codes: 2am9 2ama 2amb
16645434 N.Sharifi, and W.L.Farrar (2006).
Androgen receptor as a therapeutic target for androgen independent prostate cancer.
  Am J Ther, 13, 166-170.  
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.