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Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes.
Peptides. 2007 Jun; 28(6):1292-8.P

Abstract

Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs.

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Authors+Show Affiliations

Silva FB
Embrapa Recursos Genéticos e Biotecnologia, Brasília-DF 70770-900, Brazil.
Monteiro AC
No affiliation info available
Del Sarto RP
No affiliation info available
Marra BM
No affiliation info available
Dias SC
No affiliation info available
Figueira EL
No affiliation info available
Oliveira GR
No affiliation info available
Rocha TL
No affiliation info available
Souza DS
No affiliation info available
da Silva MC
No affiliation info available
Franco OL
No affiliation info available
Grossi-de-Sa MF
No affiliation info available

MeSH

Amino Acid SequenceAnimalsBase SequenceBinding SitesCloning, MolecularColeopteraConserved SequenceCysteine EndopeptidasesCysteine Proteinase InhibitorsDNA, ComplementaryEscherichia coliMolecular Sequence DataProtein BindingProtein Sorting SignalsProtein Structure, TertiaryRecombinant ProteinsSequence Homology, Amino AcidSubstrate SpecificityThioredoxins

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17485144

Citation

Silva, F B., et al. "Proregion of Acanthoscelides Obtectus Cysteine Proteinase: a Novel Peptide With Enhanced Selectivity Toward Endogenous Enzymes." Peptides, vol. 28, no. 6, 2007, pp. 1292-8.
Silva FB, Monteiro AC, Del Sarto RP, et al. Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. Peptides. 2007;28(6):1292-8.
Silva, F. B., Monteiro, A. C., Del Sarto, R. P., Marra, B. M., Dias, S. C., Figueira, E. L., Oliveira, G. R., Rocha, T. L., Souza, D. S., da Silva, M. C., Franco, O. L., & Grossi-de-Sa, M. F. (2007). Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. Peptides, 28(6), 1292-8.
Silva FB, et al. Proregion of Acanthoscelides Obtectus Cysteine Proteinase: a Novel Peptide With Enhanced Selectivity Toward Endogenous Enzymes. Peptides. 2007;28(6):1292-8. PubMed PMID: 17485144.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. AU - Silva,F B, AU - Monteiro,A C S, AU - Del Sarto,R P, AU - Marra,B M, AU - Dias,S C, AU - Figueira,E L Z, AU - Oliveira,G R, AU - Rocha,T L, AU - Souza,D S L, AU - da Silva,M C M, AU - Franco,O L, AU - Grossi-de-Sa,M F, Y1 - 2007/03/31/ PY - 2007/01/23/received PY - 2007/03/21/revised PY - 2007/03/22/accepted PY - 2007/5/9/pubmed PY - 2007/9/6/medline PY - 2007/5/9/entrez SP - 1292 EP - 8 JF - Peptides JO - Peptides VL - 28 IS - 6 N2 - Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs. SN - 0196-9781 UR - https://www.unboundmedicine.com/medline/citation/17485144/Proregion_of_Acanthoscelides_obtectus_cysteine_proteinase:_a_novel_peptide_with_enhanced_selectivity_toward_endogenous_enzymes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0196-9781(07)00109-X DB - PRIME DP - Unbound Medicine ER -