Abstract
A 28-kDa fragment (residues 178-423) of the human retinoic acid receptor gamma, hRARgamma D3E, encompassing the ligand-binding domain (LBD) was overproduced in Escherichia coli and purified as a monomer to more than 95% purity and homogeneity. The Kd for all-trans retinoic acid binding was 0.6 +/- 0.1 nM. Crystals of the LBD complexed with all-trans retinoic acid were grown at pH 7 from sodium acetate in the presence of detergents using the vapor diffusion method. They diffract to 2.0 A using a synchrotron radiation (lambda=0.91 A) and belong to the tetragonal space group P4(1)2(1)2 with unit cell parameters a=b=60.6 A and c=155.3 A, one monomer per asymmetric unit, a solvent content of ca. 33%, and a Vm value of approximately 2 A3/dalton.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Cloning, Molecular
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Crystallization
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Crystallography, X-Ray
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DNA Primers
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli
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Humans
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Kinetics
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Peptide Fragments / chemistry*
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Peptide Fragments / isolation & purification
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Peptide Fragments / metabolism
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Polymerase Chain Reaction
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Receptors, Retinoic Acid / chemistry*
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Receptors, Retinoic Acid / isolation & purification
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Receptors, Retinoic Acid / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Retinoic Acid Receptor gamma
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Sequence Tagged Sites
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Tretinoin / chemistry*
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Tretinoin / metabolism*
Substances
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DNA Primers
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Peptide Fragments
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Receptors, Retinoic Acid
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Recombinant Proteins
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Tretinoin