title: Overproduction of full-length and truncated human estrogen receptors in Escherichia coli.
abstract: The full-length human estrogen receptor (hER) as well as two overlapping peptides of hER were overproduced in Escherichia coli JM109 cells, using the inducible pIC vector system. The N-terminal receptor peptide contains the DNA-binding domain as well as the hinge region, whereas the C-terminal peptide contains the same hinge region and the hormone-binding domain. Typically, 1-6 mg of estrogen receptor (ER) peptides can be recovered from 1 L E. coli cell cultures. The majority of the overexpressed proteins are found in inclusion bodies, which allow the isolation of ER peptides in high yields and of 50-80% purity. Induction for short time periods at 10 microM inducer yielded up to 50% of the ER peptides in soluble form with full biological activity. Both the intact receptor and the C-terminal fragment specifically bound estrogens and antiestrogens, whereas ER peptides that contained the DNA-binding domain were retained on a DNA-agarose resin.
authors: McArdle Laboratory for Cancer Research, University of Wisconsin-Madison 53706.
authors: H
authors: H
authors: Ahrens
authors: T J
authors: TJ
authors: Schuh
authors: B L
authors: BL
authors: Rainish
authors: J D
authors: JD
authors: Furlow
authors: J
authors: J
authors: Gorski
authors: G C
authors: GC
authors: Mueller
mesh_terms: D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D004958:Estradiol / Q000378:metabolism; D004965:Estrogen Antagonists / Q000378:metabolism; D004967:Estrogens / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007700:Kinetics; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011960:Receptors, Estrogen / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D004965:Estrogen Antagonists; D004967:Estrogens; D010446:Peptide Fragments; D011960:Receptors, Estrogen; D011994:Recombinant Proteins; D004958:Estradiol; D004247:DNA