PMID: 9183021May 1, 1997Paper

Analysis of the large aqueous pores produced by a Bacillus thuringiensis protein insecticide in Manduca sexta midgut-brush-border-membrane vesicles

European Journal of Biochemistry
J Carroll, D J Ellar

Abstract

An osmotic swelling assay utilising carboxyfluorescein self-quenching to measure intravesicular volume changes was adapted to investigate permeability changes induced by the Bacillus thuringiensis Cry1Ac delta-endotoxin in Manduca sexta midgut-brush-border-membrane vesicles (BBMV). This assay provides a more quantitative analysis of Cry-toxin-induced BBMV permeability changes, extending our previously published protocol which employed a light-scattering signal to monitor delta-endotoxin activity [Carroll, J. & Ellar, D. J. (1993) Eur. J. Biochem. 214, 771-778]. The fluorescence signal changes, supported by electron microscopy of the BBMV, demonstrated that Cry1Ac altered the membrane permeability for large non-electrolyte solutes. With this approach Cry1Ac was observed to induce or form pores freely permeant for raffinose (1.14 nm diameter) and using non-electrolytes of increasing size the pores were estimated to have a limiting diameter of approximately 2.4-2.6 nm under alkaline pH conditions.

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Citations

Oct 19, 2011·The Journal of Biological Chemistry·Nicolas GroulxRikard Blunck
Sep 6, 2003·Applied and Environmental Microbiology·Mohd Amir F AbdullahDonald H Dean
Oct 7, 2004·Applied and Environmental Microbiology·Vincent VachonLuke Masson
Feb 26, 2000·In Vitro Cellular & Developmental Biology. Animal·M J LoebE A Clark
Aug 15, 2006·Biophysical Journal·Martin KirouacRaynald Laprade
Jul 2, 1999·Applied and Environmental Microbiology·C RangR Laprade
Sep 4, 2017·Biochimica Et Biophysica Acta. Biomembranes·Thittaya KunthicPanadda Boonserm
Dec 19, 2002·Biochimica Et Biophysica Acta·Olivier PeyronnetJean-Louis Schwartz
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·E SchnepfD H Dean

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