TY - JOUR
T1 - Histochemical analysis of Bacillus thuringiensis CrylA toxin binding to midgut epithelial cells of Bombyx mori
AU - Hossain, Delwar M.
AU - Hayakawa, Tohru
AU - Shitomi, Yasuyuki
AU - Itoh, Kimiko
AU - Mitsui, Toshiaki
AU - Sato, Ryoichi
AU - Hori, Hidetaka
N1 - Funding Information:
This research was partially supported by research grants (12558069 and 13306006 to H.H., and 16790035 to T.H.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. A grant for Promotion of Niigata University Research Projects (T.H. and H.H.) also partially contributed to this research. H.M.D. was supported by a Jin-nai Foundation scholarship.
PY - 2007/1
Y1 - 2007/1
N2 - We analyzed the binding of the Bacillus thuringiensis insecticidal toxins, CrylAa, CrylAb and CrylAc, to midgut tissue of the silkworm, Bombyx mori with ligand blot analysis and histochemical observations. CrylAa, CrylAb and CrylAc bound to unique sets of proteins in various subcellular fractions prepared by centrifugation. CrylAa bound to various proteins in all subcellular fractions, whereas CrylAb bound to a single protein of ∼180 kDa in all fractions as shown by Western blot analysis. Cry1Ac bound to proteins which were primarily ∼100-120 kDa in all fractions. CrylA toxins were labeled with fluorescent dye and Cy3-labeled CrylAa, CrylAb and CrylAc were shown to localize primarily to the apical membrane region. However, they also localized to basement or basolateral membranes. The distribution of a 252-kDa membrane protein (P252) of the B. mori midgut, which was recently identified as a plausible candidate for receptor of CrylA toxins were also examined with histochemical methods. Substantial signals of FITC-labeled antibody against P252, even though not all, were evident in the apical cells, and these were coincident with Cy3-CrylAa and Cy3-CrylAc signals.
AB - We analyzed the binding of the Bacillus thuringiensis insecticidal toxins, CrylAa, CrylAb and CrylAc, to midgut tissue of the silkworm, Bombyx mori with ligand blot analysis and histochemical observations. CrylAa, CrylAb and CrylAc bound to unique sets of proteins in various subcellular fractions prepared by centrifugation. CrylAa bound to various proteins in all subcellular fractions, whereas CrylAb bound to a single protein of ∼180 kDa in all fractions as shown by Western blot analysis. Cry1Ac bound to proteins which were primarily ∼100-120 kDa in all fractions. CrylA toxins were labeled with fluorescent dye and Cy3-labeled CrylAa, CrylAb and CrylAc were shown to localize primarily to the apical membrane region. However, they also localized to basement or basolateral membranes. The distribution of a 252-kDa membrane protein (P252) of the B. mori midgut, which was recently identified as a plausible candidate for receptor of CrylA toxins were also examined with histochemical methods. Substantial signals of FITC-labeled antibody against P252, even though not all, were evident in the apical cells, and these were coincident with Cy3-CrylAa and Cy3-CrylAc signals.
KW - Bacillus thuringiensis
KW - Bombyx mori midgut
KW - Cy3-labeled CrylA
KW - Fluorescent microscope
KW - Histochemistry
KW - Insecticidal proteins
UR - http://www.scopus.com/inward/record.url?scp=33750715415&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33750715415&partnerID=8YFLogxK
U2 - 10.1016/j.pestbp.2006.01.011
DO - 10.1016/j.pestbp.2006.01.011
M3 - Article
AN - SCOPUS:33750715415
SN - 0048-3575
VL - 87
SP - 30
EP - 38
JO - Pesticide Biochemistry and Physiology
JF - Pesticide Biochemistry and Physiology
IS - 1
ER -