Resolution and properties of the proteinases in the larva of the mosquito, Aedes aegypti

Kunz, Peter A.

doi:10.1016/0020-1790(78)90009-4

Cited by 39 publications

(20 citation statements)

References 27 publications

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“…This conclusion is consistent with studies on similar enzymes produced by the fiddler crab (Eisen et at. 1973) and by larvae of other dipterans; the warble fly Hypoderma linea tum (Boulard and Garrone 1978;Lecroisey et al 1979;Lecroisey et al 1980), the mosquito Aedes aegypti (Kunz 1978) and the Hessian fly Mayetiola destructor (Shukle et al 1985). In addition, there was little effect of EDTA or Ca 2 + and Mg2+ ions at the concentrations studied on the activity of the collagenase.…”

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confidence: 84%

Characterization of Proteolytic and Collagenolytic Enzymes from the Larvae of Lucilia cuprina, the Sheep Blowfly

Bowles¹,

Carnegie²,

Sandeman³

1988

Aust. Jnl. Of Bio. Sci.

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Isoelectric focusing was used to characterize proteolytic enzymes in homogenate and excretory-secretory preparations of the larvae of L. cuprina, the sheep blowfly. Zymogram overlays showed that the larvae produce a number of highly active proteases which have a wide range of isoelectric points and molecular weights. The alkaline and neutral pI proteases were inhibited by phenylmethyl-sulfonylfluoride, leupeptin and aprotinin; this indicated the presence of serine in the active site. Pepstatin and the metal chelating agent ethylenediaminetetraacetic acid had no effect Oil the activity of any of the proteases. Optimal pH for activity of the proteases was between 7 and 8. In addition, the proteases were found to be heat labile. Digestion of collagen fibrils confirmed the existence of collagenolytic activity in the excretory-secretory enzyme preparations. It is suggested that these enzymes may be involved in the nutrition of the larvae and in the pathogenesis of the lesion on the skin.

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confidence: 84%

Characterization of Proteolytic and Collagenolytic Enzymes from the Larvae of Lucilia cuprina, the Sheep Blowfly

Bowles¹,

Carnegie²,

Sandeman³

1988

Aust. Jnl. Of Bio. Sci.

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“…The guts of mosquito larvae contain chymotrypsin-and trypsinlike proteases (5,16). Larvae which ingest the crystal rapidly degrade the 51-kDa protein with a transient accumulation of a 44-kDa intermediate and slowly degrade the 42-kDa protein to a protein of 39 kDa (1,4,7,8).…”

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confidence: 99%

Construction by site-directed mutagenesis of a 39-kilodalton mosquitocidal protein similar to the larva-processed toxin of Bacillus sphaericus 2362

et al. 1990

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After ingestion of the parasporal crystals of Bacillus sphaericus, mosquito larvae process the 42-kilodalton (kDa) toxin to a protein of 39 kDa, which has an increased toxicity (A. H. Broadwell and P. Baumann, Appl. Environ. Microbiol. 53:1333-1337, 1987). A similar activation is performed by trypsin and chymotrypsin. Using site-directed mutagenesis, we have constructed derivatives of the 42-kDa toxin with a deletion of 10 amino acids at the N terminus and deletions of 7, 17, or 20 amino acids at the C terminus. Toxicity for mosquito larvae was retained upon deletion of 7 or 17 amino acids but was lost upon deletion of 20 amino acids. Evidence is presented indicating that the protein containing deletions of 10 amino acids at the N terminus and 17 amino acids at the C terminus (corresponding to potential chymotrypsin cleavage sites) is similar to the 39-kDa protein produced in mosquito larvae or by digestion with chymotrypsin. Digestion with trypsin appears to generate a protein lacking 16 or 19 amino acids from the N terminus and 7 amino acids from the C terminus. As is the case with the recombinant-made 42-kDa protein, toxicity of its derivatives is dependent on the presence of a 51-kDa protein which is a component of the parasporal crystal of B. sphaericus 2362.

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“…Mosquito larvae contain chymotrypsin-and trypsinlike enzymes in their guts (5,15). Following ingestion, the 51-and 42-kDa proteins are degraded to proteins of about 43 and 39 kDa, respectively (1,7).…”

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confidence: 99%

Deletion analysis of the 51-kilodalton protein of the Bacillus sphaericus 2362 binary mosquitocidal toxin: construction of derivatives equivalent to the larva-processed toxin

Clark

Baumann

1990

J Bacteriol

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Bacillus sphaericus 2362 produces a binary toxin consisting of 51-and 42-kDa proteins, both of which are required for toxicity to mosquito larvae. Upon ingestion by larvae, these proteins are processed to 43 and 39 kDa, respectively. Using site-directed mutagenesis, we have obtained N-and C-terminal deletions of the 51-kDa protein and expressed them in B. subtilis by using the subtilisin promoter. Removal of 21 amino acids from the N terminus and 53 amino acids from the C terminus resulted in a protein with the same electrophoretic properties as the 43-kDa degradation product which accumulates in the guts of mosquito larvae. This protein was toxic only in the presence of the 42-kDa protein. A deletion of 32 amino acids at the N terminus combined with a 53-amino-acid deletion at the C terminus resulted in a protein which retained toxicity. Toxicity was lost upon a further deletion of amino acids at potential chymotrypsin sites (41 at the N terminus, 61 at the C terminus). Comparison of the processing of the 51-and the 42-kDa proteins indicated that in spite of their sequence similarity proteolysis occurred at different sites.

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Resolution and properties of the proteinases in the larva of the mosquito, Aedes aegypti

Cited by 39 publications

References 27 publications

Characterization of Proteolytic and Collagenolytic Enzymes from the Larvae of Lucilia cuprina, the Sheep Blowfly

Characterization of Proteolytic and Collagenolytic Enzymes from the Larvae of Lucilia cuprina, the Sheep Blowfly

Construction by site-directed mutagenesis of a 39-kilodalton mosquitocidal protein similar to the larva-processed toxin of Bacillus sphaericus 2362

Deletion analysis of the 51-kilodalton protein of the Bacillus sphaericus 2362 binary mosquitocidal toxin: construction of derivatives equivalent to the larva-processed toxin

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