The use of narrow leafed lupin -NLL (Lupinus angustifolius L.) as a new food is resulting in an increasing number of allergic reactions cases, particularly in atopic patients with other pre-existing legume allergies.In the current study, we have performed an extensive in silico analysis of the NLL seed β-conglutin proteins, a new family of major allergen proteins identified in NLL, and a comparison to other relevant food allergens such as peanut Ara h 1. We analysed the variability of surface residues involved in conformational IgE-binding epitopes, lineal B-and T-cell epitopes, and changes in 2-D structural elements and 3D motives, with the aim to investigate cross-allergenicity among lupin, peanut, and other different legumes. Our results revealed that considerable structural differences exist, particularly affecting 2-D elements (loops and coils), and numerous micro-heterogeneities are present in fundamental residues directly involved in epitopes differential variability. Thus, variability of residues involved in IgE-binding epitopes might be a major contributor to the observed differences in cross-allergenicity among legumes.
Insulin resistance (IR) is the main contributor to the development of type 2 diabetes. In this study, we have purified recombinant β-conglutin proteins (rβ1 to rβ4, and rβ6) from narrow-leafed lupin (NLL) by using affinity chromatography. The objective of this study was to evaluate the capacity of these β-conglutins to improve the IR state using ex vivo and in vitro systems. rβ1, rβ3, and rβ6 produced lower levels of pro-inflammatory mediator nitric oxide (about -7-fold in all cases), up-regulated mRNA expression levels of IRS-1 (+201, +173, +192%) and Glut-4 (+286, +121, +147%), increased levels of p85-PI3K (+188, +187, +137-fold) and Glut-4 (+503, +548, +515-fold) proteins, higher phosphorylation levels of the insulin signalling pathway activator p-IRS-1 and downstream mediators such as p-Akt, p-Cbl, and p-caveolin, and improved glucose uptake in insulin resistant (IR-C) culture cells. β-conglutin proteins were able to suppress the oxidative stress produced by insulin-induced resistance on PANC-1 control (C) cells by strongly reducing the protein oxidative carbonylation induced by ROS and balancing the metabolic homeostasis in IR-C cells through regulation of mRNA expression. At the same time, β-conglutins are able to reduce the levels of the pro-inflammatory mediator nitric oxide and promote the anti-oxidative capacity of cells by increasing the levels of reduced glutathione. These results suggest NLL β-conglutins might play a fundamental role as functional food components, since β-conglutins' nutraceutical properties could enhance the effectiveness of dietary improvement of type 2 diabetes complications.
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