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Overexpression, purification, and crystal structure of native ER alpha LBD.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2001 Jul; Vol. 22 (2), pp. 165-73. - Publication Year :
- 2001
-
Abstract
- Several crystal structures of human estrogen receptor alpha ligand-binding domain (hERalpha LBD) complexed with agonist or antagonist molecules have previously been solved. The proteins had been modified in cysteine residues (carboxymethylation) or renatured in urea to circumvent aggregation and denaturation problems. In this work, high-level protein expression and purification together with crystallization screening procedure yielded high amounts of soluble protein without renaturation or modifications steps. The native protein crystallizes in the space group P3(2) 21 with three molecules in the asymmetric unit. The overall structure is very similar to that previously reported for the hERalpha LBD with cysteine carboxymethylated residues thus validating the modification approach. The present strategy can be adapted to other cases where the solubility and the proper folding is a difficulty.<br /> (Copyright 2001 Academic Press.)
- Subjects :
- Cloning, Molecular
Computer Simulation
Crystallization
Crystallography, X-Ray
Dimerization
Estrogen Receptor alpha
Humans
Ligands
Models, Molecular
Peptide Fragments genetics
Peptide Fragments metabolism
Protein Folding
Protein Structure, Secondary genetics
Protein Structure, Tertiary genetics
Receptors, Estrogen genetics
Receptors, Estrogen metabolism
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins isolation & purification
Recombinant Fusion Proteins metabolism
Peptide Fragments chemistry
Peptide Fragments isolation & purification
Receptors, Estrogen chemistry
Receptors, Estrogen isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1046-5928
- Volume :
- 22
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 11437591
- Full Text :
- https://doi.org/10.1006/prep.2001.1409