Ihara H, Uemura T, Masuhara M, Ikawa S, Sugimoto K, Wadano A, Himeno M
College of Integrated Arts and Sciences, Osaka Prefecture University, Japan.
Comp Biochem Physiol B Biochem Mol Biol. 1998 May;120(1):197-204. doi: 10.1016/s0305-0491(98)10009-3.
The binding protein for Bacillus thuringiensis delta-endotoxin, CryIAa, from the brush border membrane of the midgut of Bombyx mori was purified by the dot blot method and delta-endotoxin affinity chromatography. The binding protein was purified to 235-fold enrichment from cholic acid extracts of brush border membranes from B. mori midgut by activated CryIAa-affinity chromatography and DEAE ion-exchange chromatography. The purified binding protein showed a single band of 180 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and this band specifically reacted to 125I-labeled CryIAa on Immobilon membrane. The affinity of the binding protein for CryIAa was equivalent to that of the brush border membrane vesicles and solubilized membrane proteins. Partial amino acid sequences of the binding protein showed sequence similarity to the cadherin-like binding protein for CryIAb from Manduca sexta, but not for CryIAc binding protein from M. sexta and Heliothis virescens.
通过斑点印迹法和δ-内毒素亲和层析，从家蚕中肠刷状缘膜中纯化了苏云金芽孢杆菌δ-内毒素CryIAa的结合蛋白。通过活化的CryIAa亲和层析和DEAE离子交换层析，从家蚕中肠刷状缘膜的胆酸提取物中纯化结合蛋白，使其富集了235倍。经十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析，纯化的结合蛋白呈现一条180 kDa的单带，并且该条带在Immobilon膜上与125I标记CryIAa发生特异性反应。该结合蛋白对CryIAa的亲和力与刷状缘膜囊泡和溶解的膜蛋白相当。结合蛋白的部分氨基酸序列与烟草天蛾中CryIAb的钙黏蛋白样结合蛋白具有序列相似性，但与烟草天蛾和棉铃虫中CryIAc结合蛋白无序列相似性。
