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Domain organization ofBacillus thuringiensis CryIIIA δ-endotoxin studied by denaturation in guanidine hydrochloride solutions and limited proteolysis

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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

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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

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