Hostname: page-component-84b7d79bbc-c654p Total loading time: 0 Render date: 2024-07-28T19:41:37.864Z Has data issue: false hasContentIssue false
  • English
  • Français

Genetically-Engineered Protease-Activated Triggers in a Pore-Forming Protein

Published online by Cambridge University Press:  15 February 2011

Barbara Walker ,
Nathan Walsh  and
Hagan Bayley
Barbara Walker
Affiliation:
Worcester Foundation for Experimental Biology 222 Maple Avenue, Shrewsbury, MA 01545
Nathan Walsh
Affiliation:
Worcester Foundation for Experimental Biology 222 Maple Avenue, Shrewsbury, MA 01545
Hagan Bayley
Affiliation:
Worcester Foundation for Experimental Biology 222 Maple Avenue, Shrewsbury, MA 01545
Get access

Abstract

Protease-activated triggers have been introduced Into a pore-forming protein, staphylococcal a-hemolysin (αHL). The hemolysin was remodeled by genetic engineering to form two-chain constructs with redundant polypeptide sequences at the central loop, the Integrity of which Is crucial for efficient pore formation. The new hemolysins are activated when the polypeptide extensions are removed by proteases. By alterating the protease recognition sequence in the loop, selective activation by specified proteases can be obtained. Protease-triggered pore-forming proteins might be used for the selective destruction of cancer cells that bear tumor-associated proteases. When certain two-chain constructs are treated with proteases, a full-length polypeptide chain forms as the result of a protease-mediated transpeptidation reaction. This reaction might be used to produce chimeric hemolysins that are Inaccessible by conventional routes.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 330: Symposium S – Biomolecular Materials by Design , 1993 , 209
Copyright
Copyright © Materials Research Society 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

1. Walker, B.J. and Bayley, H., Protein Eng. 7, in press (1994).CrossRefGoogle Scholar
2. Walker, B., Kaslanowicz, J., Krishnasastry, M. and Bayley, H., Protein Eng., submitted.Google Scholar
3. Krlshnasastry, M., Walker, B., Zorn, L., Kasianowicz, J. and Bayley, H. Genetically engineered pores as components of synthetic microstructures. In: Synthetic microstructures in biological research, edited by Schnur, J. and Peckerar, M. New York: Plenum, 1992, p. 41–51.CrossRefGoogle Scholar
4. Bayley, H., J. Cell. Biochem, in press (1994).Google Scholar
5. Kasianowicz, J.J., Walker, B., Krishnasastry, M. and Bayley, H., MRS Symp. Proc. 330, this volume (1994).CrossRefGoogle Scholar
6. Bhakdi, S. and Tranum-Jensen, J., Microbiol. Rev. 55, 733 (1991).CrossRefGoogle Scholar
7. Bhakdl, S., FOssle, R. and Tranum-Jensen, J., Proc. Natl. Acad. Sci. USA 78,5475 (1981).CrossRefGoogle Scholar
8. Menestrina, G., J. Membrane Biol. 90, 177 (1986).CrossRefGoogle Scholar
9. Füssle, R., Bhakdi, S., Sziegoleit, A., Tranum-Jensen, J., Kranz, T. and Wellensiek, H.-J., J. Cell Biol. 91, 83 (1981).CrossRefGoogle Scholar
10. Tobkes, N., Wallace, B.A. and Bayley, H., Biochemistry 24, 1915 (1985).CrossRefGoogle Scholar
11. Walker, B.J., Krishnasastry, M., Zorn, L. and Bayley, H., J. Biol. Chem. 267, 21782 (1992).CrossRefGoogle Scholar
12. Bayley, H., Krishnasastry, M., Walker, B. and Kasianowicz, J., MRS Symp. Proc. 292,243 (1993).CrossRefGoogle Scholar
13. Bayley, H. Mechanism of pore formation by staphylococcal a-hemolysin. In: Membrane protein transport, Vol 2, edited by Rothman, S. Greenwich, CT: JAI Press, 1994, in press.Google Scholar
14. Walker, B.J., Krlshnasastry, M. and Bayley, H., J. Biol. Chem. 268, 5285 (1993).CrossRefGoogle Scholar
15. Langer, R., Science 249, 1527 (1990).CrossRefGoogle Scholar
16. Cairns, J., Cancer Cells 1, 1 (1989).Google Scholar
17. Beardsley, T., Scl. Am. 270 #1 (January), 130 (1994).CrossRefGoogle Scholar
18. Blood, C.H. and Zetter, B.R., Biochim. Biophys. Acta 1032,89 (1990).Google Scholar
19. Pastan, I., Chaudhary, V. and FitzGerald, D.J., Ann. Rev. Biochem. 61,331 (1992).CrossRefGoogle Scholar
20. Llotta, L.A., Steeg, P.S. and Stetler-Stevenson, W.G., Cell 64, 327 (1991).CrossRefGoogle Scholar
21. Rothschild, K.J., Argade, P.V., Earnest, T.N., Huang, K.-S., London, E., Liao, M.-J., Bayley, H., Khorana, H.G. and Herzfeld, J., J. Biol. Chem. 257, 8592 (1982).CrossRefGoogle Scholar
22. Popot, J.L., Engelman, D.M., Gurel, O. and Zaccai, G., J. Mol. Biol. 210,829 (1989).CrossRefGoogle Scholar
23. Huang, K.-S., Liao, M.-J., Gupta, C.M., Royal, N., Biemann, K. and Khorana, H.G., J. Biol. Chem. 257, 8596 (1982).CrossRefGoogle Scholar