Identification and analysis of multivalent proteolytically resistant peptides from gluten: Implications for Celiac Sprue

University of Oslo, Kristiania (historical), Oslo, Norway
Journal of Proteome Research (Impact Factor: 5). 08/2005; 4(5):1732-41. DOI: 10.1021/pr050173t
Source: PubMed

ABSTRACT Dietary gluten proteins from wheat, rye, and barley are the primary triggers for the immuno-pathogenesis of Celiac Sprue, a widespread immune disease of the small intestine. Recent molecular and structural analyses of representative gluten proteins, most notably alpha- and gamma-gliadin proteins from wheat, have improved our understanding of these pathogenic mechanisms. In particular, based on the properties of a 33-mer peptide, generated from alpha-gliadin under physiological conditions, a link between digestive resistance and inflammatory character of gluten has been proposed. Here, we report three lines of investigation in support of this hypothesis. First, biochemical and immunological analysis of deletion mutants of alpha-2 gliadin confirmed that the DQ2 restricted T cell response to the alpha-2 gliadin are directed toward the epitopes clustered within the 33-mer. Second, proteolytic analysis of a representative gamma-gliadin led to the identification of another multivalent 26-mer peptide that was also resistant to further gastric, pancreatic and intestinal brush border degradation, and was a good substrate of human transglutaminase 2 (TG2). Analogous to the 33-mer, the synthetic 26-mer peptide displayed markedly enhanced T cell antigenicity compared to monovalent control peptides. Finally, in silico analysis of the gluten proteome led to the identification of at least 60 putative peptides that share the common characteristics of the 33-mer and the 26-mer peptides. Together, these results highlight the pivotal role of physiologically generated, proteolytically stable, TG2-reactive, multivalent peptides in the immune response to dietary gluten in Celiac Sprue patients. Prolyl endopeptidase treatment was shown to abolish the antigenicity of both the 33-mer and the 26-mer peptides, and was also predicted to have comparable effects on other proline-rich putatively immunotoxic peptides identified from other polypeptides within the gluten proteome.

Download full-text


Available from: Øyvind Molberg, Aug 20, 2015
  • Source
    • "Sequence of 300 amino acids usually forms hordeins or gliadins however, it is known that 8e17 amino acid residues peptides can still demonstrate toxicity for celiac patients (Shan et al., 2005; Tye- Din et al., 2010). Malting and fermentation processes alter the barley/wheat prolamins into short peptide fragments and amino acids (Colgrave et al., 2012; Comino et al., 2013) with immunization properties and hinder their further separation. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Big brewing companies often look for sustainable resource and energy management rather than for minimization of negative social impact or maximization of positive ones. That’s why sustainable degrowth in production sounds like nightmare to them. Several activities on market such as demand for product personalization forces manufacturers’ response. But few niche markets with huge personalization potential are still underestimated by big producers. To bridge the gap between company profits and sustainable de-growth, the useful tools for niche market value estimation, which could help in investments decision making, has been worked out. The medical data reports were used to assess the socio-demographic characteristic of customer of gluten-free beer. This market is still in its starting phase as big producers are not especially interested in beer products, which require significant investments in production process. A celiac disease (CD) is chronic inflammatory enteropathy induced with presence of gluten which prevalence is estimated is 1:100, what suggest that 1 person per 100 absolutely can't drink typical barley/wheat beer. The equations for calculation model were elaborated basing on medical reports of CD occurrence in Poland and then in Europe; total population and adult population data in 2013 (Eurostat); the Bart-Haas report of beer production in 2013 and average prices of gluten-free beers manufactured and sold in Europe. The final formulas of beer production increase and the value of gluten-free beer market have been elaborated. Results reveal that almost 6.5 million of adult people in Europe must comply with gluten-free diet. Value of gluten-free beer market in Europe is estimated at 3.3 billion EUR / annual for average price 3 EUR for 0, 5 l. Introduction of gluten-free beer into market will increase production by at least 0.1 % up to even 1%. It is a value in range of from 0.55 million to 5.5 million hectolitres per year.
    Journal of Cleaner Production 07/2015; DOI:10.1016/j.jclepro.2015.07.014 · 3.84 Impact Factor
  • Source
    • "This method revealed the presence of a 33-mer peptide that is highly resistant to gastrointestinal digestion, deamidated at multiple Gln residues by TG2, and immunogenic toward most celiac patient biopsy-derived intestinal T cell lines when presented on DQ2 + antigen-presenting cells (Figure 2) (Shan et al., 2002). Similar mock digestion of another representative gluten protein, γ5-gliadin, revealed a 26-mer peptide with these same properties (Shan et al., 2005b). As such, simulated gastrointestinal digestion of gluten is an effective method for identifying disease-relevant peptide substrates for preclinical testing of glutenases. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Celiac sprue is an inflammatory disease of the small intestine caused by dietary gluten and treated by adherence to a life-long gluten-free diet. The recent identification of immunodominant gluten peptides, the discovery of their cogent properties, and the elucidation of the mechanisms by which they engender immunopathology in genetically susceptible individuals have advanced our understanding of the molecular pathogenesis of this complex disease, enabling the rational design of new therapeutic strategies. The most clinically advanced of these is oral enzyme therapy, in which enzymes capable of proteolyzing gluten (i.e., glutenases) are delivered to the alimentary tract of a celiac sprue patient to detoxify ingested gluten in situ. In this chapter, we discuss the key challenges for discovery and preclinical development of oral enzyme therapies for celiac sprue. Methods for lead identification, assay development, gram-scale production and formulation, and lead optimization for next-generation proteases are described and critically assessed.
    Methods in enzymology 01/2012; 502:241-71. DOI:10.1016/B978-0-12-416039-2.00013-6 · 2.19 Impact Factor
  • Source
    • "Among the different gluten epitopes that have been identified, the a-gliadin epitopes are considered the most immunogenic (Arentz-Hansen et al., 2000a,b, 2002; Camarca et al., 2009; Janatuinen et al., 2002; Maiuri et al., 2003; Molberg et al., 2003; Schuppan et al., 2003; Vader et al., 2002) of which the Glia-a9 is a major immunodominant epitope. The Glia-a9 epitope sequence (aI) is part of the proteolytic resistant 33-mer identified by Shan et al. (2002, 2005) that has a high T-cell stimulatory effect. The 33-mer sequence (LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF) is only present in D-genome protein sequences of a/b-gliadins (Salentijn, personal communication). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Celiac disease is a T-cell mediated immune response in the small intestine of genetically susceptible individuals caused by ingested gluten proteins from wheat, rye, and barley. In the allohexaploid bread wheat (Triticum aestivum), gluten proteins are encoded by multigene loci present on the homoeologous chromosomes 1 and 6 of the three homoeologous genomes A, B, and D. The effect of deleting individual gluten loci was analyzed in a set of deletion lines of T. aestivum cv. Chinese Spring with regard to the level of T-cell stimulatory epitopes (Glia-α9 and Glia-α20) and to technological properties of the dough including mixing, stress relaxation, and extensibility.Deletion of loci encoding ω-gliadins, γ-gliadins, and LMW-glutenins located on the short arm of chromosome 1D, reduced the number of T-cell stimulatory epitopes and caused minor deterioration of dough quality by increase of elasticity. Deletion of loci encoding α-gliadins located on the short arm of chromosome 6D, resulted in a significant decrease in T-cell stimulatory epitopes. In parallel, the dough became more stiff and less elastic, which is an improvement for ‘Chinese Spring’ dough.We demonstrated that α-gliadins from wheat can largely be compensated by addition of avenins from oat to the flour to meet technological requirements.
    Journal of Cereal Science 03/2011; 53(2):206-216. DOI:10.1016/j.jcs.2010.12.004 · 1.94 Impact Factor
Show more