Bacterial expression of a natively folded extracellular domain fusion protein of the hFSH receptor in the cytoplasm of Escherichia coli

Leslie Lobel, Susan Pollak, Brandie Lustbader, Jeffrey Klein, Joyce W. Lustbader

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

We have expressed the extracellular domain of the hFSH receptor as a fusion protein with thioredoxin in the cytoplasm of an Escherichia coli strain that contains mutations in both the thioredoxin reductase and the glutathione reductase genes. The chimeric protein isolated following induction of expression was purified in a soluble form and binds hFSH with an affinity approximating that of native receptor. This truncated form of the receptor displays the same specificity as intact receptor and does not bind hCG. The protein is expressed at levels that exceed 5 mg/L in the bacterial cytoplasm. Expression of the properly folded extracellular domain of the hFSH receptor in the cytoplasm of E. coli allows the facile and economical purification of large quantities of material. This will facilitate the determination of the structure of the hormone-binding domain of this glycoprotein receptor as well as the production of epitope-specific antibodies.

Original languageEnglish
Pages (from-to)124-133
Number of pages10
JournalProtein Expression and Purification
Volume25
Issue number1
DOIs
StatePublished - 1 Jan 2002
Externally publishedYes

Keywords

  • Bacterial expression
  • Follicle-stimulating hormone
  • Fusion protein
  • hFSH receptor

ASJC Scopus subject areas

  • Biotechnology

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