Abstract
Denaturation ofBacillus thuringiensis CryIIIAδ-endotoxin—an insecticidal protein, active againstColeoptera larvae—in concentrated guanidine hydrochloride solutions was pursued by fluorescence and circular dichroism spectroscopy and limited proteolysis. It was found that the protein consists of two fragments that differ by their stability to denaturation by guanidine hydrochloride atpH 3. The less stable fragment corresponds to the N-terminalα-helical domain limited by Leu-279; the more stable one starts with Ile-280, contains about 330 amino acid residues, and corresponds to the molecule C-terminal moiety that consist of its twoβ-structural domains forming a superdomain.
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Abbreviations
- BT:
-
Bacillus thuringiensis
- Gdn-HCl:
-
guanidine hydrochloride
- PAGE:
-
electrophoresis in polyacrylamide gel
- SDS:
-
sodium dodecylsulfate
- CD:
-
circular dichroism
References
Chestukhina, G. G., Kostina, L. I., Zalunin, I. A., Kotova, T. S., Katrukha, S. P., Kuznetsov, Yu. S., and Stepanov, V. M. (1977).Biokhimiya 42, 1660–1667.
Chestukhina, G. G., Kostina, L. I., Mikhailova, A. L., Tyurin, S. A., Klepikova, F. S., and Stepanov, V. M. (1982).Arch. Microbiol. 132, 159–162.
Chestukhina, G. G., Tyurin, S. A., Kostina, L. L., Osterman, A. L., Zalunin, I. A., Khodova, O. M., and Stepanov, V. M. (1990).J. Protein Chem. 9, 501–507.
Convents, D., Houssier, C., Lasters, I., and Lauwereys, M. (1990).J. Biol. Chem. 265, 1369–1374.
Convents, D., Cherlet, M., van Damme, J., Lasters, I., and Lauwereys, M. (1991).Eur. J. Biochem. 195, 631–635.
Hodgman, T. C., and Ellar, D. J. (1990).J. DNA Sequence 1, 97–106.
Hoefte, H., and Whiteley, H. R. (1989).Microbiol. Rev. 53, 242–255.
Hoefte, H., Seurinck, J., Van Houtven, A., and Vaeck, M. (1987).Nucleic Acids Res. 15, 7183.
Krieg, A., Schnetter, W., Huger, A. M., and Langenbruch, G. A. (1987).System. Appl. Microbiol 9, 138–141.
Kronman, M. J., and Holmes, L. G. (1971).Photochem. Photobiol. 14, 113–134.
Laemmli, U. K. (1970).Nature 227, 680–685.
Li, J., Carroll, J., and Ellar, D. J. (1991).Nature 353, 815–821.
Sanchis, V., Lereclus, D., Menou, G., Chaufaux, J., Guo, S., and Lecadet, M.-M. (1989).Mol. Microbiol. 3, 229–238.
Tyurin, S. A., Chestukhina, G. G., Osterman, A. L., Khodova, O. M., Timokhina, E. A., Klepikova, F. S., and Stepanov, V. M. (1987).Biokhimiya 52, 918–926.
Zinchenko, A. A., Roomsch, L. D., and Antonov, V. K. (1976).Bioorg. Khim. 2, 803–810.
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Ort, P., Zalunin, I.A., Gasparov, V.S. et al. Domain organization ofBacillus thuringiensis CryIIIA δ-endotoxin studied by denaturation in guanidine hydrochloride solutions and limited proteolysis. J Protein Chem 14, 241–249 (1995). https://doi.org/10.1007/BF01886765
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DOI: https://doi.org/10.1007/BF01886765