Research article summary (published 18 Oct 2009):

Anopheles gambiae alkaline phosphatase is a functional receptor of Bacillus thuringiensis jegathesan Cry11Ba toxin.

Full Abstract

Alkaline phosphatases (ALPs, EC 3.1.3.1) isolated from lepidopteran and dipteran species are identified as receptors for Cry1Ac and Cry11Aa toxins, respectively [Jurat-Fuentes, J. L., and Adang, M. J. (2004) Eur. J. Biochem. 7, 3127-3135; Fernandez, L. E., et al. (2006) Biochem. J. 396, 77-84]. In our study, an alkaline phosphatase cDNA (AgALP1) was cloned from the midgut of Anopheles gambiae larvae. The encoded 63 kDa protein has a predicted glycosylphosphatidylinositol (GPI) anchor omega-site ((526)Asp), an N-glycosylation site ((239)Asn-Leu-Thr), and an O-glycosylation site ((312)Ser). AgALP1(t) was expressed in Escherichia coli and used to prepare antiserum and to analyze the interaction of AgALP with mosquitocidal Cry11Ba toxin. Anti-AgALP serum localized AgALP to the apical brush border in the anterior and posterior midgut of larvae and detected a 65 kDa species on a blot of brush border membrane vesicles (BBMVs) protein prepared from larvae. ALP activity was released from larval BBMVs prepared by phosphatidylinositol-specific phospholipase C (PIPLC) treatment, and after separation by two-dimensional gel electrophoresis and blotting, a chain of doublet spots at 65 kDa was detected by anti-AgALP. A subset of these doublet spots bound Cry11Ba on a reprobed blot. Heterologously expressed AgALP1(t) bound [(125)I]Cry11Ba on dot blots and reduced the level of binding of [(125)I]Cry11Ba to brush border membrane vesicles by 41%, a percentage comparable to that of unlabeled Cry11Ba and aminopeptidase AgAPN2(t1) peptide. AgALP1(t) binds Cry11Ba toxin with a high affinity (23.9 nM) and shares a binding site on Cry11Ba with AgAPN2(t1). In bioassays against An. gambiae larvae, the presence of AgALP1(t) reduced larval mortality from 78 to 8%. We conclude that AgALP1 is a binding protein and a functional receptor for Cry11Ba toxin.

Author information

Author/s: Hua, Gang (G); Zhang, Rui (R); Bayyareddy, Krishnareddy (K); Adang, Michael J (MJ);

Affiliation: Department of Entomology, University of Georgia,Athens, Georgia 30602-2603, USA.

Grants: R01 AI 29092 (Agency:NIAID NIH HHS)

Journal and publication information

Publication Type: Journal Article; Research Support, N.I.H., Extramural

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

Reference: 2009-Oct; vol 48 (issue 41) : pp 9785-93

Dates: Created 2009/10/13; Completed 2009/11/02;

PMID: 19747003, status: MEDLINE (last retrieval date: 11/2/2009, 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.

Alkaline Phosphatase - metabolism Amino Acid Sequence Aminopeptidases - metabolism Animals Anopheles gambiae - enzymology; genetics Bacillus thuringiensis - chemistry; genetics; metabolism Bacterial Proteins - chemistry; genetics; metabolism Bacterial Toxins - chemistry; genetics; metabolism Binding Sites Cloning, Molecular

Computational Biology Conserved Sequence DNA Primers Diptera Endotoxins - chemistry; genetics; metabolism Hemolysin Proteins - chemistry; genetics; metabolism Lepidoptera Molecular Sequence Data Pest Control, Biological Polymerase Chain Reaction

Associated Chemicals: Bacterial Proteins (0) ; Bacterial Toxins (0) ; CRY11BB protein, Bacillus thuringiensis (0) ; DNA Primers (0) ; Endotoxins (0) ; Hemolysin Proteins (0) ; insecticidal crystal protein, Bacillus Thuringiensis (0) ; Alkaline Phosphatase (EC 3.1.3.1) ; Aminopeptidases (EC 3.4.11.-)

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