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PDBsum entry 2ocf

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protein ligands Protein-protein interface(s) links
Hormone/growth factor PDB id
2ocf
Jmol
Contents
Protein chains
233 a.a. *
93 a.a. *
Ligands
EST
Waters ×11
* Residue conservation analysis
PDB id:
2ocf
Name: Hormone/growth factor
Title: Human estrogen receptor alpha ligand-binding domain in compl estradiol and the e2#23 fn3 monobody
Structure: Estrogen receptor. Chain: a. Fragment: ligand binding domain, residues 298-595. Synonym: er, estradiol receptor, er-alpha. Engineered: yes. Mutation: yes. Fibronectin. Chain: d. Fragment: e2#23 fn3 monobody, fibronectin type iii domain 1
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: esr1, esr, nr3a1. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Gene: cig, fn1, fn. Expressed in: escherichia coli. Expression_system_taxid: 562
Resolution:
2.95Å     R-factor:   0.194     R-free:   0.251
Authors: S.S.Rajan,S.M.Kuruvilla,S.K.Sharma,Y.Kim,J.Huang,A.Koide,S.K A.Joachimiak,G.L.Greene
Key ref: A.Koide et al. (2002). Probing protein conformational changes in living cells by using designer binding proteins: application to the estrogen receptor. Proc Natl Acad Sci U S A, 99, 1253-1258. PubMed id: 11818562
Date:
20-Dec-06     Release date:   06-Nov-07    
PROCHECK
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 Headers
 References

Protein chain
Pfam   ArchSchema ?
P03372  (ESR1_HUMAN) -  Estrogen receptor
Seq:
Struc:
 
Seq:
Struc:
595 a.a.
233 a.a.*
Protein chain
Pfam   ArchSchema ?
P02751  (FINC_HUMAN) -  Fibronectin
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
2386 a.a.
93 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 11 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     3 terms  

 

 
Proc Natl Acad Sci U S A 99:1253-1258 (2002)
PubMed id: 11818562  
 
 
Probing protein conformational changes in living cells by using designer binding proteins: application to the estrogen receptor.
A.Koide, S.Abbatiello, L.Rothgery, S.Koide.
 
  ABSTRACT  
 
A challenge in understanding the mechanism of protein function in biology is to establish the correlation between functional form in the intracellular environment and high-resolution structures obtained with in vitro techniques. Here we present a strategy to probe conformational changes of proteins inside cells. Our method involves: (i) engineering binding proteins to different conformations of a target protein, and (ii) using them to sense changes in the surface property of the target in cells. We probed ligand-induced conformational changes of the estrogen receptor alpha (ER alpha) ligand-binding domain (LBD). By using yeast two-hybrid techniques, we first performed combinatorial library screening of "monobodies" (small antibody mimics using the scaffold of a fibronectin type III domain) for clones that bind to ER alpha and then characterized their interactions with ER alpha in the nucleus, the native environment of ER alpha, in the presence of various ligands. A library using a highly flexible loop yielded monobodies that specifically recognize a particular ligand complex of ER alpha, and the pattern of monobody specificity was consistent with the structural differences found in known crystal structures of ER alpha-LBD. A more restrained loop library yielded clones that bind both agonist- and antagonist-bound ER alpha. Furthermore, we found that a deletion of the ER alpha F domain that is C-terminally adjacent to the LBD increased the crossreactivity of monobodies to the apo-ER alpha-LBD, suggesting a dynamic nature of the ER alpha-LBD conformation and a role of the F domain in restraining the LBD in an inactive conformation.