RARbeta ligand-binding domain bound to an SRC-1 co-activator peptide: purification, crystallization and preliminary X-ray diffraction analysis.

Abstract

Retinoids have demonstrated therapeutic efficacy in the treatment of acute promyelocytic leukaemia and in the chemoprevention of a large number of cancers. As the cellular signalling pathway of retinoids can be transduced by the three retinoic acid receptor (RAR) isotypes alpha, beta and gamma, the side effects of these treatments induced efforts to generate isotype-selective ligands. Despite knowledge of the crystal structures of RARalpha and RARgamma ligand-binding domains (LBDs), the rational design of such ligands has been hampered by the absence of RARbeta LBD structural data. Here, a strategy used to express a large-scale soluble fraction of the human RARbeta LBD suitable for biophysical analysis is reported, as well as a procedure for crystallizing it bound to a synthetic retinoid (TTNPB) with or without a co-activator peptide (SRC-1). Preliminary X-ray analysis revealed that both complexes crystallized in the orthorhombic space group P2(1)2(1)2(1). The unit-cell parameters are a = 47.81, b = 58.52, c = 92.83 A for the TTNPB-hRARbeta LBD crystal and a = 58.14, b = 84.07, c = 102.37 A when the SRC-1 peptide is also bound.

• Authors: Sabrina Kammerer, Pierre Germain, Ralf Flaig, Carole Peluso-Iltis, André Mitschler, Natacha Rochel, Hinrich Gronemeyer, Dino Moras
• Journal: Acta crystallographica. Section D, Biological crystallography (Acta Crystallogr D Biol Crystallogr) Vol. 60 Issue Pt 11 Pg. 2048-50 (Nov 2004) ISSN: 0907-4449 [Print] United States
• PMID: 15502323 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Ligands
• Peptide Fragments
• Receptors, Retinoic Acid
• Retinoids
• Transcription Factors
• retinoic acid receptor beta
• Histone Acetyltransferases
• NCOA1 protein, human
• Nuclear Receptor Coactivator 1

Topics

• Amino Acid Sequence
• Binding Sites
• Crystallization
• Crystallography, X-Ray
• Gene Expression
• Histone Acetyltransferases
• Humans
• Ligands
• Molecular Sequence Data
• Nuclear Receptor Coactivator 1
• Peptide Fragments (chemistry, genetics, isolation & purification, metabolism)
• Protein Structure, Tertiary
• Receptors, Retinoic Acid (chemistry, genetics, isolation & purification, metabolism)
• Retinoids (pharmacology)
• Transcription Factors (chemistry, metabolism)