Document Detail

Versatile copurification procedure for rapid isolation of homogeneous RAR-RXR heterodimers.
MedLine Citation:
PMID:  10419826     Owner:  NLM     Status:  MEDLINE    
RAR-RXR heterodimeric complexes (RARalphaDeltaAB-RXRalphaDeltaAB) bound to cognate DR5 DNA response elements were purified to apparent structural and functional homogeneity using a novel versatile immobilized metal affinity chromatography (IMAC) copurification procedure. Dynamic light scattering studies indicated that the complexes were more than 85% monodisperse. By electron microscopy the negatively stained RAR-RXR-DNA complexes appeared homogeneous and corresponded to a dimeric arrangement of the molecules. Using heterodimers purified according to this procedure we demonstrate ligand binding of RXR in the context of the RAR-RXR heterodimer-DNA complex. The present copurification procedure is rapid and has yielded high quality heterodimer-DNA complexes suitable for both structural and biochemical studies.
J Iyer; D Bonnier; F Granger; C Iltis; V Andry; P Schultz; H Gronemeyer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Protein expression and purification     Volume:  16     ISSN:  1046-5928     ISO Abbreviation:  Protein Expr. Purif.     Publication Date:  1999 Jul 
Date Detail:
Created Date:  1999-08-18     Completed Date:  1999-08-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  308-14     Citation Subset:  IM    
Copyright Information:
Copyright 1999 Academic Press.
Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC)/CNRS/INSERM/ULP, C.U. de Strasbourg, Illkirch Cedex, 67404, France.
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MeSH Terms
Chromatography, Affinity
Electrophoresis, Polyacrylamide Gel
Microscopy, Electron
Receptors, Retinoic Acid / chemistry,  isolation & purification*,  metabolism
Recombinant Proteins / chemistry,  isolation & purification,  metabolism
Retinoid X Receptors
Transcription Factors / chemistry,  isolation & purification*,  metabolism
Reg. No./Substance:
0/Ligands; 0/Receptors, Retinoic Acid; 0/Recombinant Proteins; 0/Retinoid X Receptors; 0/Transcription Factors

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