Skip to main content
SpringerLink
Log in

The properties of mnemiopsin, a bioluminescent and light sensitive protein purified by hollow fiber techniques

  • General Articles
  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Summary

A calcium activated photoprotein, termed mnemiopsin, which emits bioluminescence upon the addition of calcium ion, has been isolated from the Ctenophore, Mnemiopsis leidyi, and purified by hollow fiber techniques. The system is similar to aequorin, from the jellyfish Aequorea, except that mnemiopsin can be light-inactivated. Separation of mnemiopsin from the dilute and large volume animal homogenate proved difficult with conventional biochemical techniques. A continuous flow process utilizing large surface area hollow fibers for filtration, concentration, and dialysis was developed which may also be applicable to the purification of other proteins. The resulting mnemiopsin concentrate, after further purification, was judged to be about 90% pure by its gel electrophoretic profile. Estimates by molecular sieve chromatography and SDS gel electrophoresis gave a molecular weight of about 23,000 daltons. The calcium specificity for triggering light emission was studied by comparison of triggering with a variety of cations and anions and by investigating the effects of calcium ionophores and antagonists. The activity of mnemiopsin was characterized with respect to pH, temperature and ionic strength. The stability of mnemiopsin activity after exposure to proteases, denaturants, protein group-specific reagents, detergents, elevated temperatures and light was determined.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Hastings, J. W. and Morin, J. G., 1968. Biol. Bull. 135, 422.

    Google Scholar 

  2. Hastings, J. W. and Morin, J. G., 1969. Biol. Bull. 137, 402.

    Google Scholar 

  3. Morin, J. G. and Hastings, J. W., 1971a. J. Cell. Physiol. 77, 305.

    Google Scholar 

  4. Morin, J. G. and Hastings, J. W., 1971b. J. Cell. Physiol. 77, 313.

    Google Scholar 

  5. Johnson, F. H., 1959. J. Cell. Comp. Physiol. 49, 209.

    Google Scholar 

  6. Shimomura, O., Johnson, F. H. and Saiga, Y., 1962. J. Cell. Comp. Physiol. 59, 223.

    Google Scholar 

  7. Girsch, S. J. and Hastings, J. W., 1973. Abstracts Amer. Soc. Photobiol., Sarasota, Fla., 1.

  8. Ward, W. W. and Seliger, H. H., 1974a. Biochemistry 13, 1491.

    Google Scholar 

  9. Ward, W. W. and Seliger, H. H., 1974b. Biochemistry 13, 1500.

    Google Scholar 

  10. Ward, W. W. and Seliger, H. H., 1976. Photochem. Photobiol. 23, 351.

    Google Scholar 

  11. Hastings, J. W. and Weber, G., 1963. J. Opt. Soc. Am. 53, 1410.

    Google Scholar 

  12. Mitchell, G. and Hastings, J. W., 1971. Anal. Biochem. 39, 243.

    Google Scholar 

  13. Harvey, E. N., 1921. Biol. Bull. 41, 280.

    Google Scholar 

  14. Weber, K. and Osborn, M., 1969. J. Biol. Chem. 224, 4406.

    Google Scholar 

  15. Freeman, G. and Reynolds, G. T., 1973. Develop. Biol. 81, 61.

    Google Scholar 

  16. Johnson, F. H., 1970. Nav. Res. Rev. 16–23, Feb.

  17. Blinks, J. R., Prendergast, F. G. and Allen, D. G., 1976. Pharmacological Rev. 28, 1–93.

    Google Scholar 

  18. Davis, B. J., 1964. Ann. N.Y. Acad. Sci. 121, 404.

    Google Scholar 

  19. Shimomura, O. and Johnson, F. H., 1969. Biochemistry 8, 3991.

    Google Scholar 

  20. Blinks, J. R., Mattingly, P. H., Jewell, B. R. and Van Leeuwen, M., 1969. Fed. Proc. 28, 781.

    Google Scholar 

  21. Hastings, J. W. Mitchell, G. W., Mattingly, P. H., Blinks, J. R. and Van Leeuwen, M., 1969. Nature 222, 1047.

    Google Scholar 

  22. Kohama, Y., Shimomura, O. and Johnson, F. H., 1971. Biochemistry 10, 4199.

    Google Scholar 

  23. Hartree, E. F., 1972. Anal. Biochem. 48, 422.

    Google Scholar 

  24. Harvey, E. N., 1925. J. Gen. Physiol. 7, 331.

    Google Scholar 

  25. Kreiss, P. and Cormier, M. J., 1967. Biochim Biophys. Acta 141, 181.

    Google Scholar 

  26. Sweeney, B. M., Haxo, F. T. and Hastings, J. W., 1959. J. Gen. Physiol. 43, 285.

    Google Scholar 

  27. Bode, V. C., DeSa, R. J. and Hastings, J. W., 1963. Science 141, 913.

    Google Scholar 

  28. Tanford, C., 1961. In Physical Chemistry of Macromolecules, Wiley and Sons, New York, N.Y., p. 587.

    Google Scholar 

  29. Tanford, C. 1972. Biochemistry 11, 2192.

    Google Scholar 

  30. Baldwin, T. O., 1974. Fed. Proc. 33, 1441.

    Google Scholar 

  31. Njus, D., Baldwin, T. O. and Hastings, J. W., 1974. Anal. Biochem. 61, 280.

    Google Scholar 

  32. Means, G. and Feeney, R., 1972. In Chemical Modifications of Proteins, Holden-Day, Inc., San Francisco.

    Google Scholar 

  33. Carafoli, E. and Sacktor, B., 1972. Biochem. Biophys. Res. Commun. 49, 1498.

    Google Scholar 

  34. Caswell, A., 1972. J. Membrane Biol. 7, 345.

    Google Scholar 

  35. Caswell, A. and Pressman, B. 1972. Biochem. Biophys. Res. Commun. 49, 292.

    Google Scholar 

  36. Grizzuti, K. and Perlmann, G., 1973. Biochemistry 12, 4399.

    Google Scholar 

  37. Izutsu, K., Felton, S., Siegel, I., Yoda, W. and Chen, A., 1972. Biochem. Biophys. Res. Commun. 49, 1034.

    Google Scholar 

  38. Portzehl, H. Caldwell, P. and Ruegg, J., 1964. Biochem. Biophys. Acta, 79, 581.

    Google Scholar 

  39. Shimomura, O., Johnson, F. H. and Morise, H., 1974. Biochemistry 13, 3278.

    Google Scholar 

Download references

Author information

Author notes

  1. S. J. Girsch

    Present address: Institute of Molecular Biology, University of Oregon, 97403, Eugene, OR, USA

Authors and Affiliations

  1. The Biological Laboratories, Harvard University, 02138, Cambridge, Massachusetts, USA

    S. J. Girsch & J. W. Hastings

  2. The Marine Biological Laboratory, 02543, Woods Hole, Massachusetts, USA

    S. J. Girsch & J. W. Hastings

Authors
  1. S. J. Girsch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. W. Hastings
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Girsch, S.J., Hastings, J.W. The properties of mnemiopsin, a bioluminescent and light sensitive protein purified by hollow fiber techniques. Mol Cell Biochem 19, 113–124 (1978). https://doi.org/10.1007/BF00232600

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00232600

Keywords

Search

Navigation