Crystal structure of a novel Mid-gut procarboxypeptidase from the cotton pest Helicoverpa armigera

E. Estebanez-Perpica, A. Bayes, J. Vendrell, M.A. Jongsma, D.P. Bown, J.A. Gatehouse, W. Bode, R. Huber, F.X. Aviles, D. Reverter

    Research output: Contribution to journalArticleAcademicpeer-review

    32 Citations (Scopus)

    Abstract

    The cotton bollworm Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) is one of the most serious insect pests in Australia, India and China. The larva causes substantial economical losses to legume, fibre, cereal oilseed and vegetable crops. This pest has proven to be difficult to control by conventional means, mainly due to the development of pesticide resistance. We present here the 2.5 Å crystal structure from the novel procarboxypeptidase (PCPAHa) found in the gut extracts from H. armigera larvae, the first one reported for an insect. This metalloprotease is synthesized as a zymogen of 46.6 kDa which, upon in vitro activation with Lys-C endoproteinase, yields a pro-segment of 91 residues and an active carboxypeptidase moiety of 318 residues. Both regions show a three-dimensional structure quite similar to the corresponding structures in mammalian digestive carboxypeptidases, the most relevant structural differences being located in the loops between conserved secondary structure elements, including the primary activation site. This activation site contains the motif (Ala)5Lys at the C terminus of the helix connecting the pro- and the carboxypeptidase domains. A remarkable feature of PCPAHa is the occurrence of the same (Ala)6Lys near the C terminus of the active enzyme. The presence of Ser255 in PCPAHa instead of Ile and Asp found in the pancreatic A and B forms, respectively, enlarges the S1¿ specificity pocket and influences the substrate preferences of the enzyme. The C-terminal tail of the leech carboxypeptidase inhibitor has been modelled into the PCPAHa active site to explore the substrate preferences and the enzymatic mechanism of this enzyme
    Original languageEnglish
    Pages (from-to)629-638
    JournalJournal of Molecular Biology
    Volume313
    Issue number3
    DOIs
    Publication statusPublished - 2001

    Keywords

    • carboxypeptidase-a
    • 3-dimensional structure
    • activation pathway
    • complex
    • overexpression
    • lepidoptera
    • inhibition
    • cloning
    • larvae

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