PMID: 19916922Nov 18, 2009Paper

Effect of entomocidal proteins from Bacillus thuringiensis on ion permeability of apical membranes of Tenebrio molitor larvae gut epithelium

Biochemistry. Biokhimii︠a︡
I M AndreevG G Chestukhina

Abstract

Effects of entomocidal Cry-type proteins, delta-endotoxins Cry3A and Cry11A produced by Bacillus thuringiensis, on ion permeability of the apical membranes of intestinal epithelium from Tenebrio molitor larvae midgut were studied. Using potential-sensitive dyes safranine O and oxonol VI and DeltapH indicator acridine orange, it was shown that placing brush border membrane vesicles (BBMV) (loaded with Mg2+ during their preparation) into a salt-free buffer medium resulted in spontaneous generation of transmembrane electric potential on the vesicular membrane (negative inside the vesicles) accompanied by acidification of the aqueous phase inside the vesicles. The generation of transmembrane ion gradients on the vesicular membrane was a result of an electrogenic efflux of Mg2+ from the vesicles as shown by abolishing of the membrane potential by such agents as MgSO4 or CaCl2 in centimolar concentrations, a highly lipophilic cation tetraphenylphosphonium, and some blockers of cell membrane Ca2+-channels in submillimolar concentrations. A passive generation of membrane potential on the vesicular membrane (but positive inside the vesicles) was also observed upon addition of centimolar concentrations of K2SO4. Addition of delta-endotox...Continue Reading

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