The orthologue to the Cpm1/Cqm1 receptor in Aedes aegypti is expressed as a midgut GPI-anchored α-glucosidase, which does not bind to the insecticidal binary toxin

Ferreira, Lígia Maria; Romão, Tatiany Patrícia; Neto, Osvaldo P. de Melo; Silva-Filha, Maria Helena Neves Lobo

doi:10.1016/j.ibmb.2010.05.007

Cited by 38 publications

(31 citation statements)

References 44 publications

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“…Alternatively, these APNs are attached to brush border membrane via another anchorage system (discussed below). In regards to the glucosidase with predicted GPI anchorage, interestingly, the glucosidase present in DRM is not the GPI-anchored glucosidase which was examined, but refuted, as a receptor to mosquitocidal Bin toxin 60 . Overall, most of the predicted GPI-anchored proteins present in Aedes DRM are glycosidases and hydrolases.…”

Section: Discussionmentioning

confidence: 99%

Proteome Analysis of Cry4Ba Toxin-interacting Aedes aegypti Lipid Rafts using geLC–MS/MS

et al. 2012

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Lipid rafts are microdomains in the plasma membrane of eukaryotic cells. Among their many functions, lipid rafts are involved in cell toxicity caused by pore forming bacterial toxins including Bacillus thuringiensis (Bt) Cry toxins. We isolated lipid rafts from brush border membrane vesicles (BBMV) of Aedes aegypti larvae as a detergent resistant membrane (DRM) fraction on density gradients. Cholesterol, aminopeptidase (APN), alkaline phosphatase (ALP) and the raft marker flotillin were preferentially partitioned into the lipid raft fraction. When mosquitocidal Cry4Ba toxin was pre-incubated with BBMV, Cry4Ba localized to lipid rafts. A proteomic approach based on one dimensional gel electrophoresis, in-gel trypsin digestion, followed by liquid chromatography-mass spectrometry (geLC-MS/MS) identified a total of 386 proteins. Of which many are typical lipid raft marker proteins including flotillins and glycosylphosphatidylinositol (GPI)-anchored proteins. Identified raft proteins were annotated in silico for functional and physicochemical characteristics. Parameters such as distribution of isoelectric point, molecular mass, and predicted post-translational modifications relevant to lipid raft proteins (GPI anchorage and myristoylation or palmitoylation) were analyzed for identified proteins in the DRM fraction. From a functional point of view, this study identified proteins implicated in Cry toxin interactions as well as membrane-associated proteins expressed in the mosquito midgut that have potential relevance to mosquito biology and vector management.

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Section: Discussionmentioning

confidence: 99%

Proteome Analysis of Cry4Ba Toxin-interacting Aedes aegypti Lipid Rafts using geLC–MS/MS

et al. 2012

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“…On the other hand, several studies have reported the occurrence of b-fructofuranosidase in lepidopteran insects (Santos and Terra, 1986;Sumida et al, 1994a;Carneiro et al, 2004;Daimon et al, 2008). Although a large variety of a-glucosidase genes have been recently identified in Diptera (Ferreira et al, 2010;Gabrisko and Janecek, 2011;Gabrisko, 2013;Zhang et al, 2013), there are only a few studies on a-glucosidase genes of Lepidoptera. All digestive enzymes in the Lepidoptera except those required for initial digestion are localized in the membrane fraction of midgut cell homogenates (Sumida et al, 1990;Ferreira et al, 1994;Ferreira, 1994, 2012).…”

Section: Introductionmentioning

confidence: 99%

A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose

Wang

Kiuchi

Katsuma

et al. 2015

Insect Biochemistry and Molecular Biology

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“…The BinB confers specificity to the Bin toxin because it recognizes and binds to GPI‐anchored midgut α‐glucosidases, Cqm1 and Agm3, characterized as specific receptors in Culex quinquefasciatus and Anopheles gambiae larvae, respectively (Romão et al , 2006; Opota et al , 2008). In Aedes aegypti , a refractory species, the Bin toxin is not able to bind to the Aam1 α‐glucosidase, despite the fact that Aam1 is the Cqm1 orthologue, displaying higher identity than Agm3, confirming the high level of specificity required for this interaction (Ferreira et al , 2010). So far, however, not much is known regarding the molecular basis for this specificity; then the goal of this study was to evaluate N‐ and C‐terminal truncations of BinB, as well as mutants containing replacements of specific amino acid motifs, in their ability to bind to Cqm1 receptors from C. quinquefasciatus , in order to better define elements that are critical for receptor recognition and binding.…”

Section: Introductionmentioning

confidence: 99%

The N-terminal third of the BinB subunit from the Bacillus sphaericus binary toxin is sufficient for its interaction with midgut receptors in Culex quinquefasciatus

Romão¹,

Neto²,

Silva-Filha³

2011

FEMS Microbiology Letters

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Heterodimeric binary (Bin) toxin, the major insecticidal protein from Bacillus sphaericus, acts on Culex quinquefasciatus larvae through specific binding to the midgut receptor Cqm1, a role mediated by its 448-amino-acid-long BinB subunit. The molecular basis for receptor recognition is not well understood and this study attempted to identify protein segments and amino acid motifs within BinB that are required for this event. First, N- and C-terminally truncated constructs were evaluated for their capacity to bind to native Cqm1 through pull-down assays. These showed that residues N33 to L158 of the subunit are required for Cqm1 binding. Nine different full-length mutants were then generated in which selected blocks of three amino acids were replaced by alanines. In new pull-down assays, two mutants, in which residues (85) IRF(87) and (147) FQF(149) were targeted, failed to bind the receptor. Competition binding assays confirmed the requirements for the N-terminal 158 residues, and the (147) FQF(149) epitope, for the mutant proteins to compete with native Bin toxin when binding to membrane fractions from the insect midgut. The data from this work rule out the involvement of C-terminal segments in receptor binding, highlighting the need for multiple elements within the protein's N-terminal third for it to occur.

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The orthologue to the Cpm1/Cqm1 receptor in Aedes aegypti is expressed as a midgut GPI-anchored α-glucosidase, which does not bind to the insecticidal binary toxin

Cited by 38 publications

References 44 publications

Proteome Analysis of Cry4Ba Toxin-interacting Aedes aegypti Lipid Rafts using geLC–MS/MS

Proteome Analysis of Cry4Ba Toxin-interacting Aedes aegypti Lipid Rafts using geLC–MS/MS

A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose

The N-terminal third of the BinB subunit from the Bacillus sphaericus binary toxin is sufficient for its interaction with midgut receptors in Culex quinquefasciatus

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