Skip to main content

Dissection of the LXXLL nuclear receptor-coactivator interaction motif using combinatorial peptide libraries: discovery of peptide antagonists of estrogen receptors alpha and beta.

Publication ,  Journal Article
Chang, CY; Norris, JD; Grøn, H; Paige, LA; Hamilton, PT; Kenan, DJ; Fowlkes, D; McDonnell, DP
Published in: Mol Cell Biol
December 1999

Recruitment of transcriptional coactivators following ligand activation is a critical step in nuclear receptor-mediated target gene expression. Upon binding an agonist, the receptor undergoes a conformational change which facilitates the formation of a specific coactivator binding pocket within the carboxyl terminus of the receptor. This permits the alpha-helical LXXLL motif within some coactivators to interact with the nuclear receptors. Until recently, the LXXLL motif was thought to function solely as a docking module; however, it now appears that sequences flanking the core motif may play a role in determining receptor selectivity. To address this issue, we used a combinatorial phage display approach to evaluate the role of flanking sequences in influencing these interactions. We sampled more than 10(8) variations of the core LXXLL motif with estradiol-activated estrogen receptor alpha (ERalpha) as a target and found three different classes of peptides. All of these peptides interacted with ERalpha in an agonist-dependent manner and disrupted ERalpha-mediated transcriptional activity when introduced into target cells. Using a series of ERalpha-mutants, we found that these three classes of peptides showed different interaction patterns from each other, suggesting that not all LXXLL motifs are the same and that receptor binding selectivity can be achieved by altering sequences flanking the LXXLL core motif. Most notable in this regard was the discovery of a peptide which, when overexpressed in cells, selectively disrupted ERbeta- but not ERalpha-mediated reporter gene expression. This novel ERbeta-specific antagonist may be useful in identifying and characterizing the ERbeta-regulated process in estradiol-responsive cells. In conclusion, using a combinatorial approach to define cofactor-receptor interactions, we have clearly been able to demonstrate that not all LXXLL motifs are functionally equivalent, a finding which suggests that it may be possible to target receptor-LXXLL interactions to develop receptor-specific antagonists.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

December 1999

Volume

19

Issue

12

Start / End Page

8226 / 8239

Location

United States

Related Subject Headings

  • Tumor Cells, Cultured
  • Transcription, Genetic
  • Recombinant Fusion Proteins
  • Receptors, Estrogen
  • Receptors, Cytoplasmic and Nuclear
  • Peptides
  • Peptide Library
  • Molecular Sequence Data
  • Ligands
  • Humans
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Chang, C. Y., Norris, J. D., Grøn, H., Paige, L. A., Hamilton, P. T., Kenan, D. J., … McDonnell, D. P. (1999). Dissection of the LXXLL nuclear receptor-coactivator interaction motif using combinatorial peptide libraries: discovery of peptide antagonists of estrogen receptors alpha and beta. Mol Cell Biol, 19(12), 8226–8239. https://doi.org/10.1128/MCB.19.12.8226
Chang, C. Y., J. D. Norris, H. Grøn, L. A. Paige, P. T. Hamilton, D. J. Kenan, D. Fowlkes, and D. P. McDonnell. “Dissection of the LXXLL nuclear receptor-coactivator interaction motif using combinatorial peptide libraries: discovery of peptide antagonists of estrogen receptors alpha and beta.Mol Cell Biol 19, no. 12 (December 1999): 8226–39. https://doi.org/10.1128/MCB.19.12.8226.
Chang, C. Y., et al. “Dissection of the LXXLL nuclear receptor-coactivator interaction motif using combinatorial peptide libraries: discovery of peptide antagonists of estrogen receptors alpha and beta.Mol Cell Biol, vol. 19, no. 12, Dec. 1999, pp. 8226–39. Pubmed, doi:10.1128/MCB.19.12.8226.
Chang CY, Norris JD, Grøn H, Paige LA, Hamilton PT, Kenan DJ, Fowlkes D, McDonnell DP. Dissection of the LXXLL nuclear receptor-coactivator interaction motif using combinatorial peptide libraries: discovery of peptide antagonists of estrogen receptors alpha and beta. Mol Cell Biol. 1999 Dec;19(12):8226–8239.

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

December 1999

Volume

19

Issue

12

Start / End Page

8226 / 8239

Location

United States

Related Subject Headings

  • Tumor Cells, Cultured
  • Transcription, Genetic
  • Recombinant Fusion Proteins
  • Receptors, Estrogen
  • Receptors, Cytoplasmic and Nuclear
  • Peptides
  • Peptide Library
  • Molecular Sequence Data
  • Ligands
  • Humans