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The Role of Bacillus thuringiensis Cry1C and Cry1E Separate Structural Domains in the Interaction with Spodoptera littoralis Gut Epithelial Cells
Year:
2004
Source of publication :
Journal of Biological Chemistry
Authors :
Keller, Menahem L.
;
.
Volume :
279
Co-Authors:
Avisar, D., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Keller, M., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel, Department of Field and Garden Crops, Volcani Center, POB 6, Bet Dagan 50250, Israel
Gazit, E., Dept. of Molec. Microbiol./Biotech., George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Prudovsky, E., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Sneh, B., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Zilberstein, A., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel, Dept. of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Facilitators :
From page:
15779
To page:
15786
(
Total pages:
8
)
Abstract:
The Bacillus thuringiensis δ-endotoxins Cry1C and Cry1E share toxicity against several important lepidopteran species. Their combined use to delay development of resistance in target insects depends on their differential interaction with the gut epithelial cells. The three structural domains and combinations of two consecutive domains of Cry1C and Cry1E were separately expressed in Escherichia coli, and their interactions with the brush border membrane vesicles (BBMV) of Cry1E-tolerant and -susceptible Spodoptera littoralis larvae were studied. About 80% reduction in binding of Cry1E and each of its separate domains to BBMV of Cry1E-tolerant larvae was observed, whereas Cry1C was toxic to all larvae and bound equally to BBMV derived from both Cry1E-tolerant and -susceptible larvae. These results suggest differential interactions of the two toxins with BBMV encompassing all three domains. Comparable binding assays performed with fluorescent Cry1C and Cry1C domain II showed that Cry1C has higher Bmax and lower Kd than Cry1C domain II and further supported the existence of toxin multisite interactions. Competitive binding assays were used to estimate the sequence of interaction events. Cry1C domain II could compete with domain III binding, whereas domain III did not interfere with domain II binding, indicating sequential interactions of domain III and then domain II with the same membrane site. No competition between domain II of Cry1C and Cry1E was observed, confirming the existence of different domain II binding sites for the two toxins. Taken together, all three domains specifically interact with the epithelial cell membrane. The folding of the three-domain toxin probably dictates the sequence of interaction events.
Note:
Related Files :
Animals
Biochemistry
cell membrane
Hemolysin Proteins
Lepidoptera
Protein interaction
Show More
Related Content
More details
DOI :
10.1074/jbc.M312597200
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
20120
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:34
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Scientific Publication
The Role of Bacillus thuringiensis Cry1C and Cry1E Separate Structural Domains in the Interaction with Spodoptera littoralis Gut Epithelial Cells
279
Avisar, D., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Keller, M., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel, Department of Field and Garden Crops, Volcani Center, POB 6, Bet Dagan 50250, Israel
Gazit, E., Dept. of Molec. Microbiol./Biotech., George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Prudovsky, E., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Sneh, B., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
Zilberstein, A., Department of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel, Dept. of Plant Sciences, George S. Wise Fac. of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
The Role of Bacillus thuringiensis Cry1C and Cry1E Separate Structural Domains in the Interaction with Spodoptera littoralis Gut Epithelial Cells
The Bacillus thuringiensis δ-endotoxins Cry1C and Cry1E share toxicity against several important lepidopteran species. Their combined use to delay development of resistance in target insects depends on their differential interaction with the gut epithelial cells. The three structural domains and combinations of two consecutive domains of Cry1C and Cry1E were separately expressed in Escherichia coli, and their interactions with the brush border membrane vesicles (BBMV) of Cry1E-tolerant and -susceptible Spodoptera littoralis larvae were studied. About 80% reduction in binding of Cry1E and each of its separate domains to BBMV of Cry1E-tolerant larvae was observed, whereas Cry1C was toxic to all larvae and bound equally to BBMV derived from both Cry1E-tolerant and -susceptible larvae. These results suggest differential interactions of the two toxins with BBMV encompassing all three domains. Comparable binding assays performed with fluorescent Cry1C and Cry1C domain II showed that Cry1C has higher Bmax and lower Kd than Cry1C domain II and further supported the existence of toxin multisite interactions. Competitive binding assays were used to estimate the sequence of interaction events. Cry1C domain II could compete with domain III binding, whereas domain III did not interfere with domain II binding, indicating sequential interactions of domain III and then domain II with the same membrane site. No competition between domain II of Cry1C and Cry1E was observed, confirming the existence of different domain II binding sites for the two toxins. Taken together, all three domains specifically interact with the epithelial cell membrane. The folding of the three-domain toxin probably dictates the sequence of interaction events.
Scientific Publication
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