Aminopeptidase N isoforms from the midgut of Bombyx mori and Plutella xylostella -- their classification and the factors that determine their binding specificity to Bacillus thuringiensis Cry1A toxin

Kazuko Nakanishi; Katsuro Yaoi; Yasushi Nagino; Hirotaka Hara; Madoka Kitami; Shogo Atsumi; Nami Miura; Ryoichi Sato

doi:10.1016/s0014-5793(02)02708-4

Aminopeptidase N isoforms from the midgut of Bombyx mori and Plutella xylostella -- their classification and the factors that determine their binding specificity to Bacillus thuringiensis Cry1A toxin

FEBS Lett. 2002 May 22;519(1-3):215-20. doi: 10.1016/s0014-5793(02)02708-4.

Authors

Kazuko Nakanishi¹, Katsuro Yaoi, Yasushi Nagino, Hirotaka Hara, Madoka Kitami, Shogo Atsumi, Nami Miura, Ryoichi Sato

Affiliation

¹ Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan.

PMID: 12023048
DOI: 10.1016/s0014-5793(02)02708-4

Abstract

Novel aminopeptidase N (APN) isoform cDNAs, BmAPN3 and PxAPN3, from the midguts of Bombyx mori and Plutella xylostella, respectively, were cloned, and a total of eight APN isoforms cloned from B. mori and P. xylostella were classified into four classes. Bacillus thuringiensis Cry1Aa and Cry1Ab toxins were found to bind to specific APN isoforms from the midguts of B. mori and P. xylostella, and binding occurred with fragments that corresponded to the BmAPN1 Cry1Aa toxin-binding region of each APN isoform. The results suggest that APN isoforms have a common toxin-binding region, and that the apparent specificity of Cry1Aa toxin binding to each intact APN isoform seen in SDS-PAGE is determined by factors such as expression level in conjunction with differences in binding affinity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aminopeptidases / classification*
Aminopeptidases / genetics*
Aminopeptidases / metabolism
Animals
Bacillus thuringiensis Toxins
Bacterial Proteins / metabolism*
Bacterial Toxins / metabolism
Binding Sites / genetics
Bombyx / enzymology
Cloning, Molecular
DNA, Complementary / genetics
Digestive System / enzymology
Electrophoresis, Polyacrylamide Gel
Endotoxins / metabolism*
Hemolysin Proteins
Immunoblotting
Insect Proteins / classification*
Insect Proteins / genetics*
Insect Proteins / metabolism
Isoenzymes / classification
Isoenzymes / genetics
Isoenzymes / metabolism
Lepidoptera / enzymology*
Molecular Sequence Data
Peptide Fragments / chemistry
Peptide Fragments / genetics
Peptide Fragments / metabolism
Phylogeny
Protein Binding / physiology
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Sequence Homology, Amino Acid
Substrate Specificity / physiology

Substances

Bacillus thuringiensis Toxins
Bacterial Proteins
Bacterial Toxins
DNA, Complementary
Endotoxins
Hemolysin Proteins
Insect Proteins
Isoenzymes
Peptide Fragments
Recombinant Fusion Proteins
insecticidal crystal protein, Bacillus Thuringiensis
Aminopeptidases