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Molecular and biochemical characterisation of a dual proteolytic system in vine weevil larvae (Otiorhynchus sulcatus Coleoptera: Curculionidae).
Insect Biochem Mol Biol. 2010 Nov; 40(11):785-91.IB

Abstract

The ability of phytophagous insects to utilise the relatively low nitrogen content of plant tissues is typically the limiting factor in their nutritional uptake. In the larval stage, the vine weevil feeds predominantly on root tissues of plants. The root tissue as a whole has low levels of free amino acids, and thus effective hydrolysis of dietary proteins is essential for survival. In contrast to previous reports the present study demonstrates through both molecular and biochemical studies the presence of proteolytic enzymes from two mechanistic classes, cysteine and serine proteases, in the gut of larval vine weevil; with the latter being the predominant form. cDNA clones encoding cathepsin B-like and serine-like sequences were isolated from a gut specific cDNA library; the cathepsin B-like clone has the Cys-His-Asn catalytic triad. However, the sequence showed the replacement of the conserved His-His sequence in the "occluding loop" region of the enzyme with Asp-His. This may result in a change to the substrate specificity. Two trypsin precursors contained evidence of a signal peptide, activation peptide, and conserved N-termini (IVGG). Other structural features included typical His, Asp, and Ser residues of the catalytic amino acid triad indicative of serine proteases, characteristic residues in the substrate-binding pocket, and four pairs of cysteine residues for disulfide bridges. The apparent abundance of the trypsin-like cDNA clones compared to the cathepsin B clones suggests that serine proteases are the predominant form, thus supporting data from the biochemical studies.

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Authors+Show Affiliations

Edwards MG
Newcastle University, School of Biology, Institute for Research on Environment and Sustainability, Devonshire Building, Newcastle upon Tyne NE1 7RU, UK. m.g.edwards@newcastle.ac.uk
Gatehouse JA
No affiliation info available
Gatehouse AM
No affiliation info available

MeSH

Amino Acid SequenceAnimalsBase SequenceCysteine ProteasesGastrointestinal TractGene LibraryLarvaMolecular Sequence DataOligonucleotide ProbesSerine ProteasesWeevils

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20709171

Citation

Edwards, M G., et al. "Molecular and Biochemical Characterisation of a Dual Proteolytic System in Vine Weevil Larvae (Otiorhynchus Sulcatus Coleoptera: Curculionidae)." Insect Biochemistry and Molecular Biology, vol. 40, no. 11, 2010, pp. 785-91.
Edwards MG, Gatehouse JA, Gatehouse AM. Molecular and biochemical characterisation of a dual proteolytic system in vine weevil larvae (Otiorhynchus sulcatus Coleoptera: Curculionidae). Insect Biochem Mol Biol. 2010;40(11):785-91.
Edwards, M. G., Gatehouse, J. A., & Gatehouse, A. M. (2010). Molecular and biochemical characterisation of a dual proteolytic system in vine weevil larvae (Otiorhynchus sulcatus Coleoptera: Curculionidae). Insect Biochemistry and Molecular Biology, 40(11), 785-91. https://doi.org/10.1016/j.ibmb.2010.07.005
Edwards MG, Gatehouse JA, Gatehouse AM. Molecular and Biochemical Characterisation of a Dual Proteolytic System in Vine Weevil Larvae (Otiorhynchus Sulcatus Coleoptera: Curculionidae). Insect Biochem Mol Biol. 2010;40(11):785-91. PubMed PMID: 20709171.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular and biochemical characterisation of a dual proteolytic system in vine weevil larvae (Otiorhynchus sulcatus Coleoptera: Curculionidae). AU - Edwards,M G, AU - Gatehouse,John A, AU - Gatehouse,A M R, Y1 - 2010/08/13/ PY - 2009/02/06/received PY - 2010/07/19/revised PY - 2010/07/27/accepted PY - 2010/8/17/entrez PY - 2010/8/17/pubmed PY - 2011/1/21/medline SP - 785 EP - 91 JF - Insect biochemistry and molecular biology JO - Insect Biochem Mol Biol VL - 40 IS - 11 N2 - The ability of phytophagous insects to utilise the relatively low nitrogen content of plant tissues is typically the limiting factor in their nutritional uptake. In the larval stage, the vine weevil feeds predominantly on root tissues of plants. The root tissue as a whole has low levels of free amino acids, and thus effective hydrolysis of dietary proteins is essential for survival. In contrast to previous reports the present study demonstrates through both molecular and biochemical studies the presence of proteolytic enzymes from two mechanistic classes, cysteine and serine proteases, in the gut of larval vine weevil; with the latter being the predominant form. cDNA clones encoding cathepsin B-like and serine-like sequences were isolated from a gut specific cDNA library; the cathepsin B-like clone has the Cys-His-Asn catalytic triad. However, the sequence showed the replacement of the conserved His-His sequence in the "occluding loop" region of the enzyme with Asp-His. This may result in a change to the substrate specificity. Two trypsin precursors contained evidence of a signal peptide, activation peptide, and conserved N-termini (IVGG). Other structural features included typical His, Asp, and Ser residues of the catalytic amino acid triad indicative of serine proteases, characteristic residues in the substrate-binding pocket, and four pairs of cysteine residues for disulfide bridges. The apparent abundance of the trypsin-like cDNA clones compared to the cathepsin B clones suggests that serine proteases are the predominant form, thus supporting data from the biochemical studies. SN - 1879-0240 UR - https://www.unboundmedicine.com/medline/citation/20709171/Molecular_and_biochemical_characterisation_of_a_dual_proteolytic_system_in_vine_weevil_larvae__Otiorhynchus_sulcatus_Coleoptera:_Curculionidae__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0965-1748(10)00160-8 DB - PRIME DP - Unbound Medicine ER -