Global proteome changes in larvae of Callosobruchus maculatus Coleoptera: Chrysomelidae:Bruchinae) following ingestion of a cysteine proteinase inhibitor

Fábio C S Nogueira, Carlos P. Silva, Daniel Alexandre, Richard I. Samuels, Emanoella L. Soares, Francisco J L Aragão, Giuseppe Palmisano, Gilberto B. Domont, Peter Roepstorff, Francisco A P Campos

Research output: Contribution to journalArticlepeer-review

Abstract

The seed-feeding beetle Callosobruchus maculatus is an important cowpea pest (Vigna unguiculata) as well as an interesting model to study insect digestive physiology. The larvae of C. maculatus rely on cysteine and aspartic peptidases to digest proteins in their diet. In this work, the global proteomic changes induced in the intestinal tract of larval C. maculatus challenged by the ingestion of cystatin, a cysteine peptidase inhibitor, was investigated by a nanoLC-MS/MS approach. The ingestion of cystatin caused a delay in the development of the larvae, but the mortality was not high, indicating that C. maculatus is able to adapt to this inhibitor. This proteomic strategy resulted in the identification of 752 and 550 protein groups in the midgut epithelia and midgut contents, respectively, and quantitative analyses allowed us to establish relative differences of the identified proteins. Ingestion of cystatin led to significant changes in the proteome of both the midgut epithelia and midgut contents. We have observed that proteins related to plant cell wall degradation, particularly the key glycoside hydrolases of the families GH5 (endo-β-1,4-mannanase) and GH 28 (polygalacturonase) were overexpressed. Conversely, α-amylases were downexpressed, indicating that an increase in hemicelluloses digestion helps the larvae to cope with the challenge of cystatin ingestion. Furthermore, a number of proteins associated with transcription/translation and antistress reactions were among the cystatin-responsive proteins, implying that a substantial rearrangement in the proteome occurred in C. maculatus exposed to the inhibitor.

Original languageEnglish
Pages (from-to)2704-2715
Number of pages12
JournalProteomics
Volume12
Issue number17
DOIs
Publication statusPublished - Aug 2012

Keywords

  • Callosobruchus maculatus
  • Egg white cystatin
  • Insect digestive enzymes
  • Peptidase inhibitors
  • Plant proteomics
  • Vigna unguiculata

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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