Response of the digestive system of Helicoverpa zea to ingestion of potato carboxypeptidase inhibitor and characterization of an uninhibited carboxypepidase B

A. Bayes, M. Rodrigues de la Vega, J. Vendrell, F.X. Aviles, M.A. Jongsma, M.J. Beekwilder

    Research output: Contribution to journalArticleAcademicpeer-review

    28 Citations (Scopus)

    Abstract

    Carboxypeptidase activity participates in the protein digestion process in the gut of lepidopteran insects, supplying free amino-acids to developing larvae. To study the role of different carboxypeptidases in lepidopteran protein digestion, the effect of potato carboxypeptidase inhibitor (PCI) on the digestive system of larvae of the pest insect Helicoverpa zea was investigated, and compared to that of Soybean Kunitz Trypsin Inhibitor. Analysis of carboxypeptidase activity in the guts showed that ingested PCI remained active in the gut, and completely inhibited the activity of carboxypeptidases A and O. Interestingly, carboxypeptidase B activity was not affected by PCI. All previously described enzymes from the same family, both from insect or mammalian origin, have been found to be very sensitive to PCI. Analysis of several lepidopteran species showed the presence of carboxypeptidase B activity resistant to PCI in most of them. The H. zea carboxypeptidase B enzyme (CPBHz) was purified from gut content by affinity chromatography. N-terminal sequence information was used to isolate its corresponding full-length cDNA, and recombinant expression of the zymogen of CPBHz in Pichia pastoris was achieved. The substrate specificity of recombinant CPBHz was tested using peptides. Unlike other CPB enzymes, the enzyme appeared to be highly selective for C-terminal lysine residues. Inhibition by PCI appeared to be pH-dependent.
    Original languageEnglish
    Pages (from-to)654-664
    JournalInsect Biochemistry and Molecular Biology
    Volume36
    Issue number8
    DOIs
    Publication statusPublished - 2006

    Fingerprint

    Digestive system
    Carboxypeptidases
    carboxypeptidases
    Digestive System
    Helicoverpa zea
    Solanum tuberosum
    Zea mays
    digestive system
    Eating
    ingestion
    potatoes
    carboxypeptidase B
    Carboxypeptidase B
    Insects
    Lepidoptera
    Enzymes
    enzymes
    Proteolysis
    Larva
    Kunitz Soybean Trypsin Inhibitor

    Keywords

    • plant proteinase-inhibitors
    • trypsin-inhibitor
    • spodoptera-exigua
    • noctuidae larvae
    • agrotis-ipsilon
    • cdna cloning
    • sequence
    • identification
    • expression
    • armigera

    Cite this

    @article{a99167aa036d43da805d37b9b14b8cc6,
    title = "Response of the digestive system of Helicoverpa zea to ingestion of potato carboxypeptidase inhibitor and characterization of an uninhibited carboxypepidase B",
    abstract = "Carboxypeptidase activity participates in the protein digestion process in the gut of lepidopteran insects, supplying free amino-acids to developing larvae. To study the role of different carboxypeptidases in lepidopteran protein digestion, the effect of potato carboxypeptidase inhibitor (PCI) on the digestive system of larvae of the pest insect Helicoverpa zea was investigated, and compared to that of Soybean Kunitz Trypsin Inhibitor. Analysis of carboxypeptidase activity in the guts showed that ingested PCI remained active in the gut, and completely inhibited the activity of carboxypeptidases A and O. Interestingly, carboxypeptidase B activity was not affected by PCI. All previously described enzymes from the same family, both from insect or mammalian origin, have been found to be very sensitive to PCI. Analysis of several lepidopteran species showed the presence of carboxypeptidase B activity resistant to PCI in most of them. The H. zea carboxypeptidase B enzyme (CPBHz) was purified from gut content by affinity chromatography. N-terminal sequence information was used to isolate its corresponding full-length cDNA, and recombinant expression of the zymogen of CPBHz in Pichia pastoris was achieved. The substrate specificity of recombinant CPBHz was tested using peptides. Unlike other CPB enzymes, the enzyme appeared to be highly selective for C-terminal lysine residues. Inhibition by PCI appeared to be pH-dependent.",
    keywords = "plant proteinase-inhibitors, trypsin-inhibitor, spodoptera-exigua, noctuidae larvae, agrotis-ipsilon, cdna cloning, sequence, identification, expression, armigera",
    author = "A. Bayes and {Rodrigues de la Vega}, M. and J. Vendrell and F.X. Aviles and M.A. Jongsma and M.J. Beekwilder",
    year = "2006",
    doi = "10.1016/j.ibmb.2006.05.010",
    language = "English",
    volume = "36",
    pages = "654--664",
    journal = "Insect Biochemistry and Molecular Biology",
    issn = "0965-1748",
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    }

    Response of the digestive system of Helicoverpa zea to ingestion of potato carboxypeptidase inhibitor and characterization of an uninhibited carboxypepidase B. / Bayes, A.; Rodrigues de la Vega, M.; Vendrell, J.; Aviles, F.X.; Jongsma, M.A.; Beekwilder, M.J.

    In: Insect Biochemistry and Molecular Biology, Vol. 36, No. 8, 2006, p. 654-664.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Response of the digestive system of Helicoverpa zea to ingestion of potato carboxypeptidase inhibitor and characterization of an uninhibited carboxypepidase B

    AU - Bayes, A.

    AU - Rodrigues de la Vega, M.

    AU - Vendrell, J.

    AU - Aviles, F.X.

    AU - Jongsma, M.A.

    AU - Beekwilder, M.J.

    PY - 2006

    Y1 - 2006

    N2 - Carboxypeptidase activity participates in the protein digestion process in the gut of lepidopteran insects, supplying free amino-acids to developing larvae. To study the role of different carboxypeptidases in lepidopteran protein digestion, the effect of potato carboxypeptidase inhibitor (PCI) on the digestive system of larvae of the pest insect Helicoverpa zea was investigated, and compared to that of Soybean Kunitz Trypsin Inhibitor. Analysis of carboxypeptidase activity in the guts showed that ingested PCI remained active in the gut, and completely inhibited the activity of carboxypeptidases A and O. Interestingly, carboxypeptidase B activity was not affected by PCI. All previously described enzymes from the same family, both from insect or mammalian origin, have been found to be very sensitive to PCI. Analysis of several lepidopteran species showed the presence of carboxypeptidase B activity resistant to PCI in most of them. The H. zea carboxypeptidase B enzyme (CPBHz) was purified from gut content by affinity chromatography. N-terminal sequence information was used to isolate its corresponding full-length cDNA, and recombinant expression of the zymogen of CPBHz in Pichia pastoris was achieved. The substrate specificity of recombinant CPBHz was tested using peptides. Unlike other CPB enzymes, the enzyme appeared to be highly selective for C-terminal lysine residues. Inhibition by PCI appeared to be pH-dependent.

    AB - Carboxypeptidase activity participates in the protein digestion process in the gut of lepidopteran insects, supplying free amino-acids to developing larvae. To study the role of different carboxypeptidases in lepidopteran protein digestion, the effect of potato carboxypeptidase inhibitor (PCI) on the digestive system of larvae of the pest insect Helicoverpa zea was investigated, and compared to that of Soybean Kunitz Trypsin Inhibitor. Analysis of carboxypeptidase activity in the guts showed that ingested PCI remained active in the gut, and completely inhibited the activity of carboxypeptidases A and O. Interestingly, carboxypeptidase B activity was not affected by PCI. All previously described enzymes from the same family, both from insect or mammalian origin, have been found to be very sensitive to PCI. Analysis of several lepidopteran species showed the presence of carboxypeptidase B activity resistant to PCI in most of them. The H. zea carboxypeptidase B enzyme (CPBHz) was purified from gut content by affinity chromatography. N-terminal sequence information was used to isolate its corresponding full-length cDNA, and recombinant expression of the zymogen of CPBHz in Pichia pastoris was achieved. The substrate specificity of recombinant CPBHz was tested using peptides. Unlike other CPB enzymes, the enzyme appeared to be highly selective for C-terminal lysine residues. Inhibition by PCI appeared to be pH-dependent.

    KW - plant proteinase-inhibitors

    KW - trypsin-inhibitor

    KW - spodoptera-exigua

    KW - noctuidae larvae

    KW - agrotis-ipsilon

    KW - cdna cloning

    KW - sequence

    KW - identification

    KW - expression

    KW - armigera

    U2 - 10.1016/j.ibmb.2006.05.010

    DO - 10.1016/j.ibmb.2006.05.010

    M3 - Article

    VL - 36

    SP - 654

    EP - 664

    JO - Insect Biochemistry and Molecular Biology

    JF - Insect Biochemistry and Molecular Biology

    SN - 0965-1748

    IS - 8

    ER -