Interaction between wheat alpha-amylase/trypsin bi-functional inhibitor and mammalian digestive enzymes: Kinetic, equilibrium and structural characterization of binding. (15th December 2016)

• Record Type: Journal Article
• Authors: Cuccioloni, Massimiliano; Mozzicafreddo, Matteo; Ali, Ishtiaq; Bonfili, Laura; Cecarini, Valentina; Eleuteri, Anna Maria; Angeletti, Mauro
• Is Part Of: Food chemistry. Volume 213(2016)
• Page Start: 571
• Page End: 578
• Publication Date: 2016-12-15
• Subjects: Bifunctional inhibitor -- Kinetics -- Thermodynamics -- Binding mode -- Wheat -- Gluten sensitivity; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2016.07.020
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: The interaction between a wheat bifunctional inhibitor and trypsin and α-amylase was studied. Wheat ATI targets the catalytic regions of both enzymes with independent binding sites. Wheat ATI inhibits trypsin and alpha-amylase with moderate-to-high potency. The inhibition could affect the onset/progression of gastrointestinal inflammatory disorders. Abstract: Alpha-amylase/trypsin bi-functional inhibitors (ATIs) are non-gluten protein components of wheat and other cereals that can hypersensitise the human gastrointestinal tract, eventually causing enteropathies in predisposed individuals. These inhibitory proteins can act both directly by targeting specific pro-inflammatory receptors, and indirectly by impairing the activity of digestive enzymes, the latter event causing the accumulation of undigested peptides with potential immunogenic properties. Herein, according to a concerted approach based on in vitro and in silico methods we characterized kinetics, equilibrium parameters and modes of binding of the complexes formed between wheat ATI and two representative mammalian digestive enzymes, namely trypsin and alpha-amylase. Interestingly, we demonstrated ATI to target both enzymes with independent binding sites and with moderately high affinity.