Brush Border Membrane aminopeptidase-N in the Midgut of the Gypsy Moth Serves as the Receptor for the CryIA(c) Delta-Endotoxin of Bacillus Thuringiensis

Insect Biochem Mol Biol. 1995 Dec;25(10):1143-51. doi: 10.1016/0965-1748(95)00050-x.

Abstract

Aminopeptidase-N (AP-N) was purified from gypsy moth (Lymantria dispar, L.) brush border membrane vesicles (BBMV) proteins by mono-Q chromatography and Superdex-75 gel filtration in the presence of the zwitterionic detergent, CHAPS, using FPLC. The purified AP-N, identified by its enzymatic activity, had an apparent size of 100 kDa, and was identified as the unique Bacillus thuringiensis insecticidal toxin, CryIA(c), binding protein. AP-N clearly displayed strong binding to CryIA(c), exhibiting little or no binding to CryIA(a) or CryIA(b), and showing no binding for the coleopteran-specific toxin, CryIIIA. Protein blots of the BBMV proteins probed with biotin-labeled and 125I-labeled insecticidal proteins revealed that CryIAc binds only to 120 kDa protein which is a slightly larger size in comparison to purified AP-N. Antibodies raised against the gypsy moth AP-N demonstrated that the purified AP-N and the 120 kDa CryIA(c) binding protein of total BBMV proteins are antigenically identical.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacillus thuringiensis / metabolism*
  • Bacterial Proteins
  • CD13 Antigens / isolation & purification
  • CD13 Antigens / metabolism*
  • Digestive System
  • Humans
  • Insect Proteins*
  • Microvilli / enzymology
  • Molecular Sequence Data
  • Moths / enzymology*
  • Receptors, Cell Surface / isolation & purification
  • Receptors, Cell Surface / metabolism*
  • Sequence Homology, Amino Acid

Substances

  • Bacterial Proteins
  • Cry toxin receptors
  • Insect Proteins
  • Receptors, Cell Surface
  • CD13 Antigens