Ser170 of Bacillus thuringiensis Cry1Ab δ-endotoxin becomesanchored in a hydrophobic moiety upon insertion of this proteininto Manduca sexta brush border membranes

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  • Ser170 of Bacillus thuringiensis Cry1Ab delta-endotoxin becomesanchored in a hydrophobic moiety upon insertion of this proteininto Manduca sexta brush border membranes
Creator
  • Alzate, Oscar
    • Affiliation: School of Medicine, Department of Cell Biology and Physiology
    • Other Affiliation: Biochemistry Department, Biophysics Program, The Davis Heart and Lung Research Institute, The Ohio State University, Columbus, 43210; OH, USA; School of Medicine, Cl 78B, # 72A-109, Universidad Pontificia Bolivariana, Medellín, Colombia
  • Hemann, Craig F
    • Other Affiliation: The Davis Heart and Lung Research Institute, The Ohio State University, Columbus, 43210; OH, USA
  • Osorio, Cristina
    • Other Affiliation: Biochemistry Department, The Ohio State University, Columbus, 43210; OH, USA
  • Hille, Russ
    • Other Affiliation: Biochemistry Department, Biophysics Program, The Ohio State University, Columbus, 43210; OH, USA; Department of Biochemistry, University of California, Riverside, 92526; CA, USA
  • Dean, Donald H
    • Other Affiliation: Biochemistry Department, Biophysics Program, The Ohio State University, Columbus, 43210; OH, USA
Abstract
  • Background Three spin-labeled mutant proteins, mutated at the beginning, middle, and end of α-helix 5 of the Bacillus thuringiensis Cry1Ab δ-endotoxin, were used to study the involvement of these specific amino acid residues in ion transport and to determine conformational changes in the vicinity of these residues when the protein was translocated into a biological membrane. Results Amino acid residue leucine 157, located in the N-terminal portion of α-helix 5, showed no involvement in ion transport, and the environment that surrounds the residue did not show any change when transferred into the biological membrane. Serine 170, located in the middle of the α-helix, showed no involvement in ion transport, but our findings indicate that in the membrane-bound state this residue faces an environment that makes the spin less mobile, as opposed to the mobility observed in an aqueous environment. Serine 176, located in the C-terminal end of the α-helix 5 is shown to be involved in ion transport activity. Conclusion Ion transport data for L157, S170, and S176, along with the mobility of the spin-labels, structural characterization of the resulting proteins, and toxicity assays against a target insect, suggest that the toxin undergoes conformational changes upon protein translocation into the midgut membrane. These conformational changes result in the midregion of the α-helix 5 being exposed to a hydrophobic-like environment. The location of these three residues in the toxin suggests that the entire α-helix becomes inserted in the insect midgut membrane.
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Resource type
  • Article
Rights statement
  • In Copyright
Rights holder
  • Oscar Alzate et al.; licensee BioMed Central Ltd.
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Journal title
  • BMC Biochemistry
Journal volume
  • 10
Journal issue
  • 1
Page start
  • 25
Language
  • English
Is the article or chapter peer-reviewed?
  • Yes
ISSN
  • 1471-2091
Bibliographic citation
  • BMC Biochemistry. 2009 Oct 19;10(1):25
Publisher
  • BioMed Central Ltd
Access right
  • Open Access
Date uploaded
  • August 23, 2012

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