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

dc.creatorRomão, Tatiany Patrícia
dc.creatorMelo-Neto, Osvaldo Pompílio de
dc.creatorSilva-Filha, Maria Helena Neves Lobo
dc.date2018-04-23T11:58:38Z
dc.date2018-04-23T11:58:38Z
dc.date2011
dc.date.accessioned2023-09-26T21:00:06Z
dc.date.available2023-09-26T21:00:06Z
dc.identifierROMÃO, T. P.; DE-MELO-NETO, O. P.; SILVA-FILHA, M. H. N. L. 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. FEMS microbiology letters, v. 321, n. 2, p. 167–174, ago. 2011.
dc.identifier1574-6968
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/26021
dc.identifier10.1111/j.1574-6968.2011.02325.x
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8867518
dc.descriptionEste estudo foi financiado pela Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco-FACEPE (outorga APQ 0427-2.13 / 08).
dc.descriptionHeterodimeric 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.
dc.description2050-01-01
dc.formatapplication/pdf
dc.languageeng
dc.rightsrestricted access
dc.subjectToxina bin
dc.subjectReceptor
dc.subjectCqm1
dc.subjectEpítopos
dc.subjectMosquito
dc.subjectBin toxin
dc.subjectReceptor
dc.subjectCqm1
dc.subjectEpitopes
dc.subjectMosquito
dc.subjectSequência de Aminoácidos
dc.subjectAnimais
dc.subjectToxinas Bacterianas / Química
dc.subjectToxinas bacterianas / metabolismo
dc.subjectLigação, competitiva
dc.subjectMembrana celular
dc.subjectCulex / metabolismo
dc.subjectEletroforese em Gel de Poliacrilamida
dc.subjectImmunoblotting
dc.subjectProteínas de Insetos / química
dc.subjectProteínas de Insetos / metabolismo
dc.subjectLarva
dc.subjectMicrovilosidades / metabolismo
dc.subjectMutagênese, Sítio-Dirigida
dc.subjectMutação
dc.subjectLigação proteica
dc.subjectSubunidades Proteicas
dc.subjectReceptores da Superfície Celular / química
dc.subjectReceptores da Superfície Celular / metabolismo
dc.subjectProteínas Recombinantes
dc.titleThe N-terminal third of the BinB subunit from the Bacillus sphaericus binary toxin is sufficient for its interaction with midgut receptors in Culex quinquefasciatus
dc.typeArticle


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