Properties of purified gut trypsin from Helicoverpa zea, adapted to to proteinase inhibitors.

M. Volpicella, L.R. Ceci, T. America, R. Gallarani, W. Bode, M.A. Jongsma, J. Beekwilder

    Research output: Contribution to journalArticleAcademicpeer-review

    95 Citations (Scopus)

    Abstract

    Pest insects such as Helicoverpa spp. frequently feed on plants expressing protease inhibitors. Apparently, their digestive system can adapt to the presence of protease inhibitors. To study this, a trypsin enzyme was purified from the gut of insects that were raised on an inhibitor-containing diet. The amino-acid sequence of this enzyme was analysed by tandem MS, which allowed assignment of 66 f the mature protein amino acid sequence. This trypsin, called HzTrypsin-S, corresponded to a known cDNA sequence from Helicoverpa. The amino acid sequence is closely related (76 dentical) to that of a trypsin, HzTrypsin-C, which was purified and identified in a similar way from insects raised on a diet without additional inhibitor. The digestive properties of HzTrypsin-S and HzTrypsin-C were compared. Both trypsins appeared to be equally efficient in degrading protein. Four typical plant inhibitors were tested in enzymatic measurements. HzTrypsin-S could not be inhibited by > 1000-fold molar excess of any of these. The same inhibitors inhibited HzTrypsin-C with apparent equilibrium dissociation constants ranging from 1 nm to 30 nm. Thus, HzTrypsin-S seems to allow the insect to overcome different defensive proteinase inhibitors in plants.
    Original languageEnglish
    Pages (from-to)10-19
    JournalEuropean Journal of Biochemistry
    Volume270
    DOIs
    Publication statusPublished - 2003

    Fingerprint

    Trypsin
    Zea mays
    Insects
    Peptide Hydrolases
    Amino Acid Sequence
    Nutrition
    Protease Inhibitors
    Amino Acids
    Digestive system
    Diet
    Digestive System
    Enzymes
    Proteins
    Complementary DNA

    Keywords

    • partial-purification
    • protease inhibitors
    • phage display
    • manduca-sexta
    • resistance
    • insects
    • plants
    • larvae
    • midgut
    • expression

    Cite this

    Volpicella, M. ; Ceci, L.R. ; America, T. ; Gallarani, R. ; Bode, W. ; Jongsma, M.A. ; Beekwilder, J. / Properties of purified gut trypsin from Helicoverpa zea, adapted to to proteinase inhibitors. In: European Journal of Biochemistry. 2003 ; Vol. 270. pp. 10-19.
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    abstract = "Pest insects such as Helicoverpa spp. frequently feed on plants expressing protease inhibitors. Apparently, their digestive system can adapt to the presence of protease inhibitors. To study this, a trypsin enzyme was purified from the gut of insects that were raised on an inhibitor-containing diet. The amino-acid sequence of this enzyme was analysed by tandem MS, which allowed assignment of 66 f the mature protein amino acid sequence. This trypsin, called HzTrypsin-S, corresponded to a known cDNA sequence from Helicoverpa. The amino acid sequence is closely related (76 dentical) to that of a trypsin, HzTrypsin-C, which was purified and identified in a similar way from insects raised on a diet without additional inhibitor. The digestive properties of HzTrypsin-S and HzTrypsin-C were compared. Both trypsins appeared to be equally efficient in degrading protein. Four typical plant inhibitors were tested in enzymatic measurements. HzTrypsin-S could not be inhibited by > 1000-fold molar excess of any of these. The same inhibitors inhibited HzTrypsin-C with apparent equilibrium dissociation constants ranging from 1 nm to 30 nm. Thus, HzTrypsin-S seems to allow the insect to overcome different defensive proteinase inhibitors in plants.",
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    author = "M. Volpicella and L.R. Ceci and T. America and R. Gallarani and W. Bode and M.A. Jongsma and J. Beekwilder",
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    Properties of purified gut trypsin from Helicoverpa zea, adapted to to proteinase inhibitors. / Volpicella, M.; Ceci, L.R.; America, T.; Gallarani, R.; Bode, W.; Jongsma, M.A.; Beekwilder, J.

    In: European Journal of Biochemistry, Vol. 270, 2003, p. 10-19.

    Research output: Contribution to journalArticleAcademicpeer-review

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    AU - Ceci, L.R.

    AU - America, T.

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    AU - Bode, W.

    AU - Jongsma, M.A.

    AU - Beekwilder, J.

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    KW - partial-purification

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    KW - phage display

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    KW - insects

    KW - plants

    KW - larvae

    KW - midgut

    KW - expression

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