Wheat Subtilisin/Chymotrypsin Inhibitor (WSCI) as a scaffold for novel serine protease inhibitors with a given specificity

Francesca Tedeschi, Antimo Di Maro, Angelo Facchiano, Susan Costantini, Angela Chambery, Natalia Bruni, Valeria Capuzzi, Anna Grazia Ficca, Elia Poerio

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

WSCI (Wheat Subtilisin/Chymotrypsin Inhibitor) is a small protein belonging to the Potato inhibitor I family exhibiting a high content of essential amino acid. In addition to bacterial subtilisins and mammalian chymotrypsins, WSCI inhibits chymotrypsin-like activities isolated from digestive traits of a number of insect larvae. In vivo, as suggested for many plant proteinase inhibitors, WSCI seems to play a role of natural defence against attacks of pests and pathogens. The functional region of WSCI, containing the inhibitor reactive site (Met48-Glu49), corresponds to an extended flexible loop (Val42-Asp53) whose architecture is somehow stabilized by a number of secondary interactions established with a small β-sheet located underneath. The aim of this study was to employ a WSCI molecule as a stable scaffold to obtain recombinant inhibitors with new acquired anti-proteinase activity or, alternatively, inactive WSCI variants. A gene sequence coding for the native WSCI, along with genes coding for muteins with different specficities, could be exploited to obtain transformed non-food use plants with improved insect resistance. On the other hand, the genetic transformation of cereal plants over-expressing inactive WSCI muteins could represent a possible strategy to improve the nutritional quality of cereal-based foods, without risk of interference with human or animal digestive enzymes. Here, we described the characterization of four muteins containing single/multiple amino acid substitutions at the WSCI reactive site and/or at its proximity. Modalities of interaction of these muteins with proteinases (subtilisin, trypsin and chymotrypsin) were investigated by time course hydrolysis and molecular simulations studies.

Original languageEnglish
Pages (from-to)3335-3343
Number of pages9
JournalMolecular BioSystems
Volume8
Issue number12
DOIs
Publication statusPublished - 2012

Fingerprint

Subtilisin
Serine Proteinase Inhibitors
Chymotrypsin
Triticum
Peptide Hydrolases
Insects
Catalytic Domain
Subtilisins
Genetic Transformation
Essential Amino Acids
Nutritive Value
Amino Acid Substitution
Solanum tuberosum
Genes
Larva
Hydrolysis

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology

Cite this

Tedeschi, F., Di Maro, A., Facchiano, A., Costantini, S., Chambery, A., Bruni, N., ... Poerio, E. (2012). Wheat Subtilisin/Chymotrypsin Inhibitor (WSCI) as a scaffold for novel serine protease inhibitors with a given specificity. Molecular BioSystems, 8(12), 3335-3343. https://doi.org/10.1039/c2mb25320h

Wheat Subtilisin/Chymotrypsin Inhibitor (WSCI) as a scaffold for novel serine protease inhibitors with a given specificity. / Tedeschi, Francesca; Di Maro, Antimo; Facchiano, Angelo; Costantini, Susan; Chambery, Angela; Bruni, Natalia; Capuzzi, Valeria; Ficca, Anna Grazia; Poerio, Elia.

In: Molecular BioSystems, Vol. 8, No. 12, 2012, p. 3335-3343.

Research output: Contribution to journalArticle

Tedeschi, F, Di Maro, A, Facchiano, A, Costantini, S, Chambery, A, Bruni, N, Capuzzi, V, Ficca, AG & Poerio, E 2012, 'Wheat Subtilisin/Chymotrypsin Inhibitor (WSCI) as a scaffold for novel serine protease inhibitors with a given specificity', Molecular BioSystems, vol. 8, no. 12, pp. 3335-3343. https://doi.org/10.1039/c2mb25320h
Tedeschi, Francesca ; Di Maro, Antimo ; Facchiano, Angelo ; Costantini, Susan ; Chambery, Angela ; Bruni, Natalia ; Capuzzi, Valeria ; Ficca, Anna Grazia ; Poerio, Elia. / Wheat Subtilisin/Chymotrypsin Inhibitor (WSCI) as a scaffold for novel serine protease inhibitors with a given specificity. In: Molecular BioSystems. 2012 ; Vol. 8, No. 12. pp. 3335-3343.
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