, Volume 14, Issue 2, pp 205–212 | Cite as

High-level bacterial expression of a natively folded, soluble extracellular domain fusion protein of the human luteinizing hormone/chorionic gonadotropin receptor in the cytoplasm of Escherichia coli

  • Leslie I. Lobel
  • Susan Pollak
  • Jeffrey Klein
  • Joyce W. Lustbader


We have expressed the extracellular domain of the human luteinizing hormone/chorionic gonadotropin (hLH/CG) receptor as a fusion protein with thioredoxin in the cytoplasm of an Escherichia coli strain that contains mutations in both the thioredoxin reductase and glutathione reductase genes. The chimeric protein isolated following induction of expression is purified in a soluble form and binds hCG with an affinity approximating that of native receptor. This truncated form of the receptor displays the same specificity as intact hLH/CG receptor and does not bind human follicle stimulating hormone. This cytoplasmically produced protein is expressed at levels that exceed 10 mg/L. Expression of properly folded extracellular domain of the hLH/CG receptor in the cytoplasm of E. coli allows the facile and economic purification of large quantities of material. This will facilitate the determination of the structure of the hormone-binding domain of the glycoprotein receptor as well as the production of epitope-specific antibodies.

Key Words

Human luteinizing hormone/chorionic gonadotropin receptor chorionic gonadotropin bacterial expression fusion protein 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Ingley, E. and Hemmings, B. A. (1999). Protein Exp. Purif. 17, 224–230.CrossRefGoogle Scholar
  2. 2.
    Qiu, J., Swartz, J. R., and Georgiou, G. (1998). Appl. Environ. Microbiol. 64, 4891–4896.PubMedGoogle Scholar
  3. 3.
    Cheadle, C., Ivashchenko, Y., South, V., Searfoss, G. H., French, S., Howk, R., Ricca, G. A., and Jaye, M. (1994). J. Biol. Chem. 269, 24,034–24,039.Google Scholar
  4. 4.
    Huth, J. R., Norton, S. E., Lockridge, O., Shikone, T., Hsueh, A. J., and Ruddon, R. W. (1994). Endocrinology 135, 911–918.PubMedCrossRefGoogle Scholar
  5. 5.
    Lobel, L. I., Rausch, P., Trakht, I., Pollak, S., and Lustbader, J. W. (1997). Endocrinology 138, 1232–1239.PubMedCrossRefGoogle Scholar
  6. 6.
    di Marzo, V., Willis, A. E., Boyer-Thompson, C., Appella, E., and Perham, R. N. (1994). J. Mol. Biol. 243, 167–172.CrossRefGoogle Scholar
  7. 7.
    Greenwood, J., Willis, A. E., and Perham, R. N. (1991). J. Mol. Biol. 220, 821–827.PubMedCrossRefGoogle Scholar
  8. 8.
    Chen, W. and Bahl, O. P. (1993). Mol. Cell. Endocrinol. 91, 35–41.PubMedCrossRefGoogle Scholar
  9. 9.
    Osuga, Y., Kudo, M., Kaipia, A., Kobilka, B., and Hsueh, A. J. (1997). Mol. Endocrinol. 11, 1659–1668.PubMedCrossRefGoogle Scholar
  10. 10.
    Prinz, W. A., Aslund, F., Holmgren, A., and Beckwith, J. (1997). J. Biol. Chem. 272, 15,661–15,667.CrossRefGoogle Scholar
  11. 11.
    Stewart, E. J., Aslund, F., and Beckwith, J. (1998). EMBO J. 17, 5543–5550.PubMedCrossRefGoogle Scholar
  12. 12.
    Wimalasena, J., Dostal, R., and Meehan, D. (1992). Gynecol. Oncol. 92, 345–350.CrossRefGoogle Scholar
  13. 13.
    Meduri, G., Charnaux, N., Loosfelt, H., Jolivet, A., Spyratos, F., Brailly, S., and Milgrom, E. (1997). Cancer Res. 57, 857–864.PubMedGoogle Scholar
  14. 14.
    Lobel, L., Pollak, S., Wang, S., Chaney, M., and Lustbader, J. W. (1999). Endocrine 10, 261–270.PubMedGoogle Scholar
  15. 15.
    Dufau, M. L. and Kusuda, S. (1987). J Recept. Res. 7, 167–193.PubMedGoogle Scholar
  16. 16.
    Dufau, M. L. (1998). Annu. Rev. Physiol. 60, 461–496.PubMedCrossRefGoogle Scholar
  17. 17.
    Lustbader, J. W., Yarmush, D. L., Birken, S., Puett, D., and Canfield, R. E. (1993). Endocr. Rev. 14, 291–311.PubMedCrossRefGoogle Scholar
  18. 18.
    O’Connor, J. F., Birken, S., Lustbader, J. W., Krichevsky, A., Chen, Y., and Canfield, R. E. (1994). Endocr. Rev. 15, 650–683.PubMedCrossRefGoogle Scholar
  19. 19.
    Cole, L. A., Shahabi, S., Oz, U. A., Bahado-Singh, R. O., and Mahoney, M. J. (1999). Clin. Chem. 45, 2109–2119.PubMedGoogle Scholar
  20. 20.
    Birken, S., Chen, Y., Gawinowicz, M. A., Lustbader, J. W., Pollak, S., Agosto, G., Buck, R., and O’Connor, J. (1993). Endocrinology 133, 1390–1397.PubMedCrossRefGoogle Scholar
  21. 21.
    Birken, S., Gawinowicz, M. A., Kardana, A., and Cole, L. A. (1991). Endocrinology 129, 1551–1558.PubMedCrossRefGoogle Scholar
  22. 22.
    Laemmli, U. K. and Favre, M. (1973). J. Mol. Biol. 73, 575–599.CrossRefGoogle Scholar
  23. 23.
    Laemmli, U. K. (1970). Nature 70, 680–685.CrossRefGoogle Scholar
  24. 24.
    Morrissey, J. H. (1981). Anal. Biochem. 117, 307–310.PubMedCrossRefGoogle Scholar
  25. 25.
    Towbin, H., Staehelin, T., and Gordon, J. (1979). Proc. Natl. Acad. Sci. USA 76, 4350–4354.PubMedCrossRefGoogle Scholar
  26. 26.
    Burnette, W. N. (1981). Anal. Biochem. 112, 195–203.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc 2001

Authors and Affiliations

  • Leslie I. Lobel
    • 1
  • Susan Pollak
    • 1
  • Jeffrey Klein
    • 1
  • Joyce W. Lustbader
    • 1
  1. 1.Center for Reproductive Science, Department of Obstetrics and GynecologyColumbia UniversityNew York

Personalised recommendations