Cargando…

Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin

Brush border membrane vesicles (BBMVs) were prepared from the 2(nd) instar larvae of Helicoverpa armigera. Binding of the activated Cry1Ac of Bacillus thuringiensis (Bt) toxin was shown by immunoblot. A 120-kDa protein was identified as a receptor for the Cry1Ac type δ-endotoxin. The aminopeptidase-...

Descripción completa

Detalles Bibliográficos
Autores principales: Ingle, Sanjay S., Trivedi, Nidhi, Prasad, Rakesh, Kuruvilla, Jes, Rao, Koteshwara Kukatla, Chhatpar, H.S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer-Verlag 2001
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7082820/
https://www.ncbi.nlm.nih.gov/pubmed/11683359
http://dx.doi.org/10.1007/s002840010297
_version_ 1783508421511741440
author Ingle, Sanjay S.
Trivedi, Nidhi
Prasad, Rakesh
Kuruvilla, Jes
Rao, Koteshwara Kukatla
Chhatpar, H.S.
author_facet Ingle, Sanjay S.
Trivedi, Nidhi
Prasad, Rakesh
Kuruvilla, Jes
Rao, Koteshwara Kukatla
Chhatpar, H.S.
author_sort Ingle, Sanjay S.
collection PubMed
description Brush border membrane vesicles (BBMVs) were prepared from the 2(nd) instar larvae of Helicoverpa armigera. Binding of the activated Cry1Ac of Bacillus thuringiensis (Bt) toxin was shown by immunoblot. A 120-kDa protein was identified as a receptor for the Cry1Ac type δ-endotoxin. The aminopeptidase-N activity of BBMVs was measured as the hydrolysis of L-leucine p-nitroanilide. The specific activity was 35 units/mg protein. The BBMV preparation also showed low level of alkaline phosphatase activity. Zn(++) chelating agents 2,2′-dipyridyl and 1,10-phenanthroline inhibited aminopeptidase activity at 10 mM concentration, indicating the presence of zinc-dependent aminopeptidase in the brush border of H. armigera. The aminopeptidase activity was increased with increasing concentration of δ-endotoxin. The purified 120-kDa binding protein was N-terminally sequenced. The first 10-amino-acid sequence showed 60–77% similarity with human cysteine-rich secretory protein-1 precursor, inhibin alpha chain precursor. Salmonella flagellar hook protein and yeast carboxypeptidase S.
format Online
Article
Text
id pubmed-7082820
institution National Center for Biotechnology Information
language English
publishDate 2001
publisher Springer-Verlag
record_format MEDLINE/PubMed
spelling pubmed-70828202020-03-23 Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin Ingle, Sanjay S. Trivedi, Nidhi Prasad, Rakesh Kuruvilla, Jes Rao, Koteshwara Kukatla Chhatpar, H.S. Curr Microbiol Article Brush border membrane vesicles (BBMVs) were prepared from the 2(nd) instar larvae of Helicoverpa armigera. Binding of the activated Cry1Ac of Bacillus thuringiensis (Bt) toxin was shown by immunoblot. A 120-kDa protein was identified as a receptor for the Cry1Ac type δ-endotoxin. The aminopeptidase-N activity of BBMVs was measured as the hydrolysis of L-leucine p-nitroanilide. The specific activity was 35 units/mg protein. The BBMV preparation also showed low level of alkaline phosphatase activity. Zn(++) chelating agents 2,2′-dipyridyl and 1,10-phenanthroline inhibited aminopeptidase activity at 10 mM concentration, indicating the presence of zinc-dependent aminopeptidase in the brush border of H. armigera. The aminopeptidase activity was increased with increasing concentration of δ-endotoxin. The purified 120-kDa binding protein was N-terminally sequenced. The first 10-amino-acid sequence showed 60–77% similarity with human cysteine-rich secretory protein-1 precursor, inhibin alpha chain precursor. Salmonella flagellar hook protein and yeast carboxypeptidase S. Springer-Verlag 2001 /pmc/articles/PMC7082820/ /pubmed/11683359 http://dx.doi.org/10.1007/s002840010297 Text en © Springer-Verlag New York Inc. 2001 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Article
Ingle, Sanjay S.
Trivedi, Nidhi
Prasad, Rakesh
Kuruvilla, Jes
Rao, Koteshwara Kukatla
Chhatpar, H.S.
Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title_full Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title_fullStr Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title_full_unstemmed Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title_short Aminopeptidase-N from the Helicoverpa armigera (Hubner) Brush Border Membrane Vesicles as a Receptor of Bacillus thuringiensis Cry1Ac δ-Endotoxin
title_sort aminopeptidase-n from the helicoverpa armigera (hubner) brush border membrane vesicles as a receptor of bacillus thuringiensis cry1ac δ-endotoxin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7082820/
https://www.ncbi.nlm.nih.gov/pubmed/11683359
http://dx.doi.org/10.1007/s002840010297
work_keys_str_mv AT inglesanjays aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin
AT trivedinidhi aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin
AT prasadrakesh aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin
AT kuruvillajes aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin
AT raokoteshwarakukatla aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin
AT chhatparhs aminopeptidasenfromthehelicoverpaarmigerahubnerbrushbordermembranevesiclesasareceptorofbacillusthuringiensiscry1acdendotoxin