Digestive enzymes in close and distant genera of a same family: Properties of midgut hydrolases from luminescent Pyrophorus divergens (Coleoptera: Elateridae) larvae

Colepicolo-Neto, P.; Bechara, Etelvino J.H.; Ferreira, Clélia; Terra, Walter R.

doi:10.1016/0305-0491(87)90384-1

Cited by 2 publications

(2 citation statements)

References 18 publications

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“…These proteinases have not been characterized further yet. The range of proteinase activities measured here in cerambycid larvae are similar to those reported in the midgut of several other coleopteran families: serine endopeptidases have been detected in larval scarabids (So0 Hoo and Dudzinski, 1967;McGhie et al, 1995), dermestids (Baker, 1981;Christeller et al, 1993), elaterids (Colepicolo-Net0 et al, 1987), carabids (Cheeseman and Gillott, 1987), tenebrionids (Levinsky et al, 19771, and curculionids (Baker, 1982). An interesting feature is the extremely high levels of leucine aminopeptidase activity in the midgut of A. gfabripennis.…”

Section: Major Proteinases In the Midgut Of A Glabvipennissupporting

confidence: 82%

Midgut proteinase activities in larvae of Anoplophora glabripennis (Coleoptera: Cerambycidae) and their interaction with proteinase inhibitors

Bian¹,

Shaw²,

Han³

et al. 1996

Arch. Insect Biochem. Physiol.

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The major proteinase activities in the larval midgut of a common poplar tree borer, Anoplophora glabripennis, were characterised. Overall digestive capacity, as measured by casein hydrolysis, showed a p H optimum between 10 and 11.5, suggestive of serine endopeptidase activity. Trypsin, chymotrypsin, and chymotrypsin-like activities were detected using specific p-nitroaniiide synthetic substrates and by use of specific serine endopeptidase inhibitors. These activities also showed pH optima in the extreme alkaline range. The absence of cysteine, aspartic, and metallo-endopeptidases were confirmed using class specific proteinase inhibitors. The dominant exopeptidase in the midgut is leucine aminopeptidase with a p H optimum of 7-9. Carboxypeptidase a and b activity were barely detectable. A large range of serine endopeptidase inhibitors were screened and were found to vary widely in their ability to inhibit casein hydrolysis. Potato proteinase inhibitor 1 (a chyrnotrypsin inhibitor) and wheatgerm trypsin inhibitor 1 inhibited particularly effectively in tandem and represent possible candidates for gene transformation to produce plants tolerant to this pest. 0 1996 Wiley-Liss, Inc.

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Section: Major Proteinases In the Midgut Of A Glabvipennissupporting

confidence: 82%

Midgut proteinase activities in larvae of Anoplophora glabripennis (Coleoptera: Cerambycidae) and their interaction with proteinase inhibitors

Bian¹,

Shaw²,

Han³

et al. 1996

Arch. Insect Biochem. Physiol.

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“…It is commonly believed that coleopteran insects depend on cysteine proteases for the majority of digestive protease activity (6,38); however, digestive serine protease activity has been identified in at least seven different families of Coleoptera (10,11,19,40,43,53,58). Such activity has also been identified in extracts from second-instar WCR larvae (17), but the contribution and role of this serine protease activity in larvae remain uncertain.…”

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confidence: 99%

An Engineered Chymotrypsin/Cathepsin G Site in Domain I Renders Bacillus thuringiensis Cry3A Active against Western Corn Rootworm Larvae

Walters

Stacy

Lee

et al. 2008

Appl Environ Microbiol

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The western corn rootworm remains one of the most important pests of corn in the United States despite the use of many pest management tools. Cry3A, the first coleopteran-active Bacillus thuringiensis toxin isolated, has not been useful for control of the corn rootworm pest complex. Modification of Cry3A so that it contained a chymotrypsin/cathepsin G protease recognition site in the loop between ␣-helix 3 and ␣-helix 4 of domain I, however, resulted in consistent activity of the toxin ("mCry3A") against neonate western corn rootworm. In vitro chymotrypsin digests showed that there was a substantial difference between the enzyme sensitivity of mCry3A and the enzyme sensitivity of Cry3A, with mCry3A rapidly converted from a 67-kDa form to a ϳ55-kDa form. The introduced protease site was also recognized in vivo, where the ϳ55-kDa form of mCry3A toxin was rapidly generated and associated with the membrane fraction. After a point mutation in mcry3A that resulted in the elimination of the native domain I chymotrypsin site (C terminal to the introduced chymotrypsin/ cathepsin G protease site of mCry3A), the in vitro and in vivo digestion patterns remained the same, demonstrating that the introduced site was required for the enhanced activity. Also, 55-kDa mCry3A generated by cleavage with chymotrypsin exhibited specific binding to western corn rootworm brush border membrane, whereas untreated 67-kDa mCry3A did not. These data indicate that the mCry3A toxicity for corn rootworm larvae was due to the introduction of a chymotrypsin/cathepsin G site, which enhanced cleavage and subsequent binding of the activated toxin to midgut cells.

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Digestive enzymes in close and distant genera of a same family: Properties of midgut hydrolases from luminescent Pyrophorus divergens (Coleoptera: Elateridae) larvae

Cited by 2 publications

References 18 publications

Midgut proteinase activities in larvae of Anoplophora glabripennis (Coleoptera: Cerambycidae) and their interaction with proteinase inhibitors

Midgut proteinase activities in larvae of Anoplophora glabripennis (Coleoptera: Cerambycidae) and their interaction with proteinase inhibitors

An Engineered Chymotrypsin/Cathepsin G Site in Domain I Renders Bacillus thuringiensis Cry3A Active against Western Corn Rootworm Larvae

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