Identification and characterization of digestive serine proteases from inhibitor-resistant Helicoverpa zea larval midgut

M. Volpicella, J.H.G. Cordewener, M.A. Jongsma, R. Gallerani, L.R. Ceci, M.J. Beekwilder

    Research output: Contribution to journalArticleAcademicpeer-review

    13 Citations (Scopus)

    Abstract

    Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence
    Original languageEnglish
    Pages (from-to)26-32
    JournalJournal of Chromatography. B, Analytical technologies in the biomedical and life sciences
    Volume833
    Issue number1
    DOIs
    Publication statusPublished - 2006

    Fingerprint

    Serine Proteinase Inhibitors
    Zea mays
    Peptide Hydrolases
    Trypsin
    Insects
    Chymotrypsin
    Nutrition
    Protease Inhibitors
    Larva
    Digestive system
    Diet
    Affinity chromatography
    Digestive System
    Affinity Chromatography
    Mass spectrometry
    Mass Spectrometry
    Enzymes
    trypsin drug combination chymotrypsin

    Keywords

    • plant proteinase-inhibitors
    • soybean trypsin-inhibitor
    • agrotis-ipsilon
    • gut proteinases
    • complex
    • expression
    • induction
    • insects
    • tobacco
    • genes

    Cite this

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    title = "Identification and characterization of digestive serine proteases from inhibitor-resistant Helicoverpa zea larval midgut",
    abstract = "Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence",
    keywords = "plant proteinase-inhibitors, soybean trypsin-inhibitor, agrotis-ipsilon, gut proteinases, complex, expression, induction, insects, tobacco, genes",
    author = "M. Volpicella and J.H.G. Cordewener and M.A. Jongsma and R. Gallerani and L.R. Ceci and M.J. Beekwilder",
    year = "2006",
    doi = "10.1016/j.jchromb.2005.10.021",
    language = "English",
    volume = "833",
    pages = "26--32",
    journal = "Journal of Chromatography. B, Analytical technologies in the biomedical and life sciences",
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    Identification and characterization of digestive serine proteases from inhibitor-resistant Helicoverpa zea larval midgut. / Volpicella, M.; Cordewener, J.H.G.; Jongsma, M.A.; Gallerani, R.; Ceci, L.R.; Beekwilder, M.J.

    In: Journal of Chromatography. B, Analytical technologies in the biomedical and life sciences, Vol. 833, No. 1, 2006, p. 26-32.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

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    AU - Volpicella, M.

    AU - Cordewener, J.H.G.

    AU - Jongsma, M.A.

    AU - Gallerani, R.

    AU - Ceci, L.R.

    AU - Beekwilder, M.J.

    PY - 2006

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    N2 - Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence

    AB - Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence

    KW - plant proteinase-inhibitors

    KW - soybean trypsin-inhibitor

    KW - agrotis-ipsilon

    KW - gut proteinases

    KW - complex

    KW - expression

    KW - induction

    KW - insects

    KW - tobacco

    KW - genes

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    DO - 10.1016/j.jchromb.2005.10.021

    M3 - Article

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