Purification and partial amino acid sequences of the binding protein from Bombyx mori for CryIAa delta-endotoxin of Bacillus thuringiensis.
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.
Ihara
H
H
College of Integrated Arts and Sciences, Osaka Prefecture University, Japan.
Uemura
T
T
Masuhara
M
M
Ikawa
S
S
Sugimoto
K
K
Wadano
A
A
Himeno
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive / Q000502:physiology; D012831:Bombyx / Q000737:chemistry*; D003001:Cloning, Molecular; D004731:Endotoxins / Q000378:metabolism*; D006460:Hemolysin Proteins; D008565:Membrane Proteins / Q000737:chemistry*; D008871:Microvilli / Q000737:chemistry; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000502:physiology; D011971:Receptors, Immunologic / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D008565:Membrane Proteins; D011971:Receptors, Immunologic; C025357:endotoxin binding proteins; C048071:endotoxin receptor; C030956:insecticidal crystal protein, Bacillus Thuringiensis