An alpha-amylase is a novel receptor for Bacillus thuringiensis ssp. israelensis Cry4Ba and Cry11Aa toxins in the malaria vector mosquito Anopheles albimanus (Diptera: Culicidae).
Bacillus thuringiensis ssp. israelensis (Bti) produces four Cry toxins (Cry4Aa, Cry4Ba, Cry10Aa and Cry11Aa), and two Cyt proteins (Cyt1Aa and Cyt2Ba), toxic to mosquito-larvae of the genus Aedes, Anopheles and Culex, important human disease vectors that transmit dengue virus, malaria and filarial parasites respectively. Previous work showed that Bti is highly toxic to Anopheles albimanus, the main vector for transmission of malaria in Mexico. In this work, we analysed the toxicity of isolated Cry proteins of Bti and identified an An. albimanus midgut protein as a putative Cry4Ba and Cry11Aa receptor molecule. Biossays showed that Cry4Ba and Cry11Aa of Bti are toxic to An. albimanus larvae. Ligand blot assays indicated that a 70 kDa glycosylphosphatidylinositol-anchored protein present in midgut brush border membrane vesicles of An. albimanus interacts with Cry4Ba and Cry11Aa toxins. This protein was identified as an alpha-amylase by mass spectrometry and enzymatic activity assays. The cDNA that codes for the alpha-amylase was cloned by means of 5'- and 3'-RACE experiments. Recombinant alpha-amylase expressed in Escherichia coli specifically binds Cry4Ba and Cry11Aa toxins.
Fernandez-Luna
Maria Teresa
MT
Depto. de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México Apdo. Postal 510-3, Cuernavaca, Morelos 62250, México.
Lanz-Mendoza
Humberto
H
Gill
Sarjeet S
SS
Bravo
Alejandra
A
Soberon
Mario
M
Miranda-Rios
Juan
J
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000033:anatomy & histology / Q000201:enzymology* / Q000382:microbiology* / Q000502:physiology; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D004731:Endotoxins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007306:Insecticides / Q000737:chemistry / Q000378:metabolism*; D007814:Larva / Q000187:drug effects / Q000502:physiology; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000516:alpha-Amylases / Q000378:metabolism*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D000516:alpha-Amylases