title: Probing the mechanism of action of Bacillus thuringiensis insecticidal proteins by site-directed mutagenesis--a minireview.
abstract: The current model of the mechanism of action of several Bacillus thuringiensis insecticidal crystal proteins (Cry) is reviewed and tested by site-directed mutagenesis experiments. Amino acid (aa) residues were substituted in each of the three domains of Cry toxins and the effects on toxin stability, binding to receptors, irreversible insertion into the membrane, and ion channel activity were examined. Mutant proteins with aa altered on the putative membrane-proximal surface of domain I are affected in insertion into the membrane and toxicity, but not in binding to the receptor. Alterations in the putative receptor-binding loops of domain II show an effect on the initial (reversible) binding to the receptor when certain aa are altered, while affecting irreversible binding when other aa are altered. Mutant proteins with aa altered in a conserved track of aa of domain III have altered ion channel properties, as measured by the voltage clamping of insect midguts and the K+ permeability of brush border membrane vesicles. In summary, domain I is involved in insertion into the membrane and affects ion channel function, domain II is involved in receptor binding and insertion into the membrane, and domain III is involved ion channel function, receptor binding, and insertion into the membrane.
authors.affiliation: Department of Biochemistry, Ohio State University, Columbus, 43210, USA. dean.10@osu.edu
authors.forename: D H
authors.initials: DH
authors.lastname: Dean
authors.forename: F
authors.initials: F
authors.lastname: Rajamohan
authors.forename: M K
authors.initials: MK
authors.lastname: Lee
authors.forename: S J
authors.initials: SJ
authors.lastname: Wu
authors.forename: X J
authors.initials: XJ
authors.lastname: Chen
authors.forename: E
authors.initials: E
authors.lastname: Alcantara
authors.forename: S R
authors.initials: SR
authors.lastname: Hussain
mesh_terms: D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000235:genetics / Q000494:pharmacology*; D004731:Endotoxins / Q000235:genetics / Q000494:pharmacology*; D006460:Hemolysin Proteins; D007306:Insecticides / Q000494:pharmacology*; D008962:Models, Theoretical; D016297:Mutagenesis, Site-Directed; D013329:Structure-Activity Relationship
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; C030956:insecticidal crystal protein, Bacillus Thuringiensis