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  • Title: Versatile copurification procedure for rapid isolation of homogeneous RAR-RXR heterodimers.
    Author: Iyer J, Bonnier D, Granger F, Iltis C, Andry V, Schultz P, Gronemeyer H.
    Journal: Protein Expr Purif; 1999 Jul; 16(2):308-14. PubMed ID: 10419826.
    Abstract:
    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.
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