The Bacillus thuringiensis delta-endotoxin. Evidence for a two domain structure of the minimal toxic fragment.
The conformational characteristics of the minimal toxic fragment of the delta-endotoxin from Bacillus thuringiensis berliner 1715 were examined by fluorescence and circular dichroism spectroscopy. This insecticidal protein, specifically toxic to lepidopteran species, was found to consist of two structural domains. Experimental evidence for this conclusion was provided by biphasic guanidine hydrochloride unfolding curves at different pH values and electrophoretic patterns of protease digests. Two stable fragments of comparable molecular weight were obtained using four different broad specificity proteolytic enzymes. A secondary structure model was constructed using seven B. thuringiensis toxin sequences. These toxins were selected on the basis of their limited sequence homology and represent all known insecticidal specificities. Despite this divergence, a consensus secondary structure pattern was obtained, confirming the structural homology among the toxins. The N-terminal halves of all toxins are predicted to be relatively rich in alpha-helix structure and the C-terminal parts to contain alternating beta-strand and coil structures. The latter seems characteristic for a beta-sheet conformation. Comparing this model to the unfolding data obtained by circular dichroism, whose far UV signal gives a measure of the alpha-helix content, allowed us to delineate the structural domains into the primary structure.
Convents
D
D
Plant Genetic Systems, Gent, Belgium.
Houssier
C
C
Lasters
I
I
Lauwereys
M
M
D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000648:ultrastructure*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000648:ultrastructure; D001427:Bacterial Toxins; D002942:Circular Dichroism; D004731:Endotoxins; D006460:Hemolysin Proteins; D010449:Peptide Mapping; D011487:Protein Conformation; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis