Research article summary (published 20 Sep 2009):

Cloning and epitope mapping of Cry11Aa-binding sites in the Cry11Aa-receptor alkaline phosphatase from Aedes aegypti.

Full Abstract

Cry11Aa is the most active Bacillus thuringiensis israelensis toxin against Aedes aegypti larvae. Ae. aegypti alkaline phosphatase (ALP) was previously identified as a Cry11Aa receptor mediating toxicity. Here we report the cloning and functional characterization of this Ae. aegypti Cry11Aa-ALP receptor. Of three ALP's cDNA clones, the recombinant produced ALP1 isoform was shown to bind Cry11Aa and P1.BBMV peptide phage that specifically binds the midgut ALP-Cry11Aa receptor. An anti-ALP1 antibody inhibited binding to brush border membrane vesicles and toxicity of Cry11Aa in isolated cultured guts. Two ALP1 Cry11Aa binding regions (R59-G102 and N257-I296) were mapped by characterizing binding of Cry11Aa to nine recombinant overlapping peptides covering the ALP1 sequence. Finally, by using a peptide spot array of Cry11Aa domain III and site-directed mutagenesis, we show that the ALP1 R59-G102 region binds Cry11Aa through domain II loop alpha-8 while ALP1 N257-I296 interacts with Cry11Aa through domain III 561RVQSQNSGNN570 located in beta18-beta19. Our results show that Cry11Aa domain II and domain III are involved in the binding with two distinct binding sites in the ALP1 receptor.

Author information

Author/s: Fernandez, Luisa E (LE); Martinez-Anaya, Claudia (C); Lira, Erandi (E); Chen, Jianwu (J); Evans, Amy (A); Hernández-Martínez, Salvador (S); Lanz-Mendoza, Humberto (H); Bravo, Alejandra (A); Gill, Sarjeet S (SS); Soberón, Mario (M);

Affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, postal 510-3, Cuernavaca 62250, Morelos, Mexico.

Grants: 1R01 AI066014 (Agency:NIAID NIH HHS)

Journal and publication information

Publication Type: Comparative Study; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

Journal: Biochemistry (Biochemistry), published in United States. (Language: eng)

Reference: 2009-Sep; vol 48 (issue 37) : pp 8899-907

Dates: Created 2009/09/16; Completed 2009/10/05;

PMID: 19697959, status: MEDLINE (last retrieval date: 10/5/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Aedes - enzymology; genetics; metabolism Alkaline Phosphatase - genetics; metabolism Amino Acid Sequence Animals Bacterial Proteins - genetics; isolation & purification; metabolism Binding Sites Cloning, Molecular Endotoxins - genetics; isolation & purification; metabolism Epitope Mapping

Epitope Mapping Hemolysin Proteins - genetics; isolation & purification; metabolism Isoenzymes - genetics; metabolism Larva - enzymology; genetics Molecular Sequence Data Mutagenesis, Site-Directed Protein Structure, Tertiary Receptors, Cell Surface - metabolism Sequence Alignment

Associated Chemicals: Bacterial Proteins (0) ; Endotoxins (0) ; Hemolysin Proteins (0) ; Isoenzymes (0) ; Receptors, Cell Surface (0) ; insecticidal crystal protein, Bacillus Thuringiensis (0) ; MC14 alkaline phosphatase I (EC 3.1.3.-) ; Alkaline Phosphatase (EC 3.1.3.1)

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