The glycoprotein toxin of Bacillus thuringiensis subsp. israelensis indicates a lectinlike receptor in the larval mosquito gut

Muthukumar, Ganapathy; Nickerson, Kenneth W.

doi:10.1128/aem.53.11.2650-2655.1987

Cited by 31 publications

(6 citation statements)

References 45 publications

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“…In contrast, when solubilized subsp. morrisoni (PG-14) crystals are incubated with gut proteases of lepidopteran larvae, a 24-kDa peptide is observed and the hemolytic activity associated with it is retained (40).…”

Section: Nc Gg/mimentioning

confidence: 99%

Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. fukuokaensis crystal proteins

Ohba

Gill

1991

Appl Environ Microbiol

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The mosquitocidal crystals of Bacillus thuringiensis subsp. fukuokaensis were isolated and bioassayed against fourth-instar larvae of two mosquito species. The 50% lethal concentration values of the crystals to Aedes aegypti and Cukx quinquefasciatus were 4.1 and 2.9 ,ug/ml, respectively. In addition, the solubilized crystals had hemolytic activity; 50 ,ug/ml was the lowest detectable level. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the crystals consisted of polypeptides of 90, 86, 82, 72, 50, 48, 37, and 27 kDa. When the solubilized inclusion was treated with C. quinquefasciatus midgut brush border membrane vesicles or Manduca sexta gut juice, only one major protein was detected. This protein retained mosquitocidal activity but had no detectable hemolytic activity. Immunological analysis of this subspecies and the subspecies israelensis, kyushuensis and darmstadiensis by using polyclonal antisera raised against the whole-crystal protein of B. thuringiensis subsp. fukuokaensis revealed that the proteins in subsp. fukuokaensis are distinct from proteins in the other subspecies because little cross-reaction was observed. Analysis of the plasmid pattern showed that the crystal protein genes are located on a plasmid of 130 MDa. Analysis of plasmid and chromosomal DNA from subsp. fukuokaensis showed little homology to the 72-kDa toxin gene (PG-14) of B. thuringiensis subsp. morisoni. However, some of the proteins of B. thuringiensis subsp. fukuokaensis are homologous to other B. thuringiensis toxins because N-terminal amino acid analysis revealed that the 90-kDa protein is encoded by a crylV gene type. * Corresponding author. kyushuensis and subsp. morrisoni PG-14 share significant homology to the crystal toxin gene of subsp. israelensis (31). More recently the monoclonal antibodies directed against the 68-and 135-kDa toxins of subsp. israelensis have been shown to cross-react with the 26-and 75or 80-kDa proteins of subsp. kyushuensis, respectively (15). In this article we describe the characterization of crystals produced from B. thuringiensis subsp. fukuokaensis which are toxic to larvae of mosquitoes and which in solubilized form are hemolytic. Further, by means of polyclonal antisera specific for crystal proteins of this strain we show that the crystal proteins of this strain do not cross-react with those of three other mosquitocidal strains. Further, we have determined that a large plasmid encodes for the crystal proteins of subsp. fukuokaensis. MATERIALS AND METHODS Bacterial strains and culture conditions. The type strains of B. thuringiensis subsp. israelensis, fukuokaensis, kyushensis, and darmstadiensis (strain 73-E10-2) were from Kyushu University, Fukuoka, Japan. Two Crymutants used in this study, subsp. israelensis YG-1 and subsp. fukuokaensis YG-101, were prepared by curing the type strains as previously described (14). Growth of these strains was as described for B. thuringiensis subsp. israelensis (10). Purification and solubilization of crystals. The spore-crysta...

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Section: Nc Gg/mimentioning

confidence: 99%

Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. fukuokaensis crystal proteins

Ohba

Gill

1991

Appl Environ Microbiol

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“…(4) The toxic and cytolytic effects of the lectinoid microbial (cholera, shiga, diphtheria, Pseudornonas, tetanus and botulinus) toxins are inhibited by sugars or glycoconjugates resembling the glycosylated membrane lipid or protein receptors of the sensitive cells [26,27]. The interesting point in this regard is the fact that Bacillus thuringiensis glycosylated toxin insecticidal activity, caused by its interaction with lectinoid receptors in the larval mosquito gut, is also inhibited by sugars [66]. (5) The plant toxins (ricin, abrin, modeccin, isidor, etc.)…”

Section: Experimental Evidence Showingmentioning

confidence: 99%

Microbial lectin cofunction with lytic activities as a model for a general basic lectin role

Gilboa‐Garber¹

1989

FEMS Microbiology Reviews

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Lectins are ubiquitous proteins, which exhibit a specific and reversible sugar-binding activity. They react with glycosylated macromolecules and cells and may coaggragate them and lead to their lysis or alterations. Various lectin biological effects are well known, but their basic biological function is considered as yet unknown. In the present review, an experimental evidence and theoretical considerations are forwarded for supporting our suggestion that the general basic lectin or lectinoid (lectin-like protein) function in microorganisms, plants and animals is a cofunction enabling the activities of key lytic enzymes (lysins: glycosidases, proteases, esterases, phosphatases, hemolysin, etc.). The lectin service is: homing onto glycosylated receptors, anchoring to them and induction of cooperative conformational effects which enable their counterpart lysin activity on exogenous or endogenous target molecules and cells. The 'lectin-lysin' pair may reside in the same molecule, or in linked subunits. It may also be formed by cofunction of two separate entities originating from one or two (homogenous or heterogenous) cell sources. The lectin and lysin may be free or cell-bound components located intra or extracellularly. The final result of their cofunction is practically irreversible; either cell and macro-molecule lysis for nutrition, homeostasis and protection or cell alteration, reorganization and new productivity. Our suggestion emphasizes the prominent analogy of lectins to lytic enzyme positioning sites (LEPS), immunoglobulins and polypeptide hormones. The lectin analogy to LEPS and immunoglobulins is exhibited in the lectin-dependent cell and macromolecule lysis for nutritional and homeostatic purposes or for protection, respectively. The hormone-like lectin activity is exhibited in the lectin-dependent cell alterations. In addition to similar functions and effects, the analogy also includes the properties and behavior of these proteins. The suggested hypothesis is based on experimental evidence from microorganisms, plants and animals. It envisions the lectin and lectinoid function in cell attacks on glycosylated molecules or cells, cell-substratum and cell-cell interactions (fusion, invasion, etc.), cell transformation and formation of special structures. All of them according to a developmental program, or special (especially unfavourable) environmental conditions. The lectin resistance to proteolysis and unfavourable pH or temperature is in accord with the suggested hypothesis.

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“…Gut agglutinins of tsetse flies, for example, are involved in controlling trypanosome infections (MAUDLIN & WELBURN, 1987). Moreover, agglutinins of the haemocoel have been demonstrated in the filariasis vectors, mosquitoes (MUTHUKUMAR & NICKERSON, 1987;Zahedi, personal communication) and blackflies . In a preliminary communication (SMAIL & HAM, 1988) these Simulium agglutinins have been shown to be lectins, but their exact function is unknown.…”

Section: Introductionmentioning

confidence: 99%

Surface carbohydrate changes onOnchocerca lienalislarvae as they develop from microfilariae to the infective third-stage inSimulium ornatum

Ham¹,

Smail²,

Groeger

1988

J. Helminthol.

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Use was made of seven FITC labelled lectins as tools to investigate the surface ofOnchocerca lienalislarvae as they develop through to the infective third-stage in a natural vector,Simulium ornatum. The lectins were derived fromCanavalia ensiformis(Con A),Lens culinaris(lentil),Triticum vulgaris(wheat germ),Arachis hypogaea(peanut),Helix pomatia, Phaseolus vulgaris(kidney bean) andTetragonolobus purpureus(asparagus pea). Between 70 and 100 living parasites were examined for each developmental stage; i.e. skin microfilariae, late first-stages, second-stages, preinfective third-stages and infective third-stages isolated from the mouth parts of the flies. None of the lectins used bound to the surface of the microfilariae. However, progressive binding to the cuticle of the first- and second-stages was observed using Con. A, lentil lectin and wheat germ agglutinin (WGA). Following moulting to the third-stage, binding of these three lectins declined. Furthermore, as these lectins decreased, peanut andHelix pomatialectins progressively increased in their binding, despite the fact that they showed little or no binding to the first- and second-stages; stages at which Con A, lentil and WGA were at their maximum. Asparagus pea and kidney bean lectins failed completely to bind to any of the larvae examined. Carbohydrate inhibition tests showed that the lectin was indeed binding specifically to glycoconjugates on the parasite surface. WGA binding was not inhibited by prior incubation with N-acetyl-D-glucosamine, even at high concentrations, but neuraminic acid did completely inhibit its binding. Judging from the patterns of binding on the nematodes themselves, the carbohydrates may not be vector in origin, but derive from the worms. The lectin specificities indicate that initially mannose/glucose type derivatives are present on the surface. Following moulting to the third-stage these are progressively replaced, or overlaid with galactosamine type derivatives, also present on the infective third-stage as it enters the bovine host. The availability of these surface glycoconjugates to attack mediated by natural insect lectins may be of importance in the parasite regulatory mechanisms of the blackfly. Variability in these surface carbohydrates, and in the response to them could well be a contributing factor in the cytospecific variation inS. damnosumsusceptibility to geographical variants ofO. volvulus.

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The glycoprotein toxin of Bacillus thuringiensis subsp. israelensis indicates a lectinlike receptor in the larval mosquito gut

Cited by 31 publications

References 45 publications

Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. fukuokaensis crystal proteins

Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. fukuokaensis crystal proteins

Microbial lectin cofunction with lytic activities as a model for a general basic lectin role

Surface carbohydrate changes onOnchocerca lienalislarvae as they develop from microfilariae to the infective third-stage inSimulium ornatum

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