Deamidation and cross-linking of gliadin peptides by transglutaminases and the relation to celiac disease.

Abstract

Activation of small intestinal gluten-reactive CD4+ T cells is a critical event in celiac disease. Such cells predominantly recognise gluten peptides in which specific glutamines are deamidated. Deamidation may be catalysed by intestinal tissue transglutaminase (TG2), a protein which is also the main autoantigen in celiac disease. Our aim was to study how the two main catalytic activities of transglutaminase--deamidation and transamidation (cross-linking) of an immunodominant gliadin epitope--are influenced by the presence of acceptor amines in the intestinal mucosa, and thereby contribute to further elucidation of the pathogenetic mechanisms in celiac disease. We prepared monoclonal antibodies, reacting specifically with the non-deamidated epitope QPFPQPQLPYPQPQ-amide and/or the deamidated epitope QPFPQPELPYPQPQ-amide. A solid phase immunoassay combined with gel filtration chromatography was used to analyse deamidation and cross-linking of these peptides to proteins. Our results show that QPFPQPQLPYPQPQ-amide was deamidated when incubated with purified TG2, with fresh mucosal sheets and with mucosal homogenates. Of other transglutaminases tested, only Streptoverticillium transglutaminase was able to generate the deamidated epitope. A fraction of the non-deamidated epitope was cross-linked to proteins, including TG2. The results suggest that intestinal TG2 is responsible for generation of the active deamidated epitope. As the epitope often occurs in a repeat structure, the result may be cross-linking of a deamidated, i.e., activated cell epitope. Alternatively, the deamidation may occur by reversal of the cross-linking reaction. The results provide a basis for the suggestion that binding of a peptide to a protein, in connection to its modification to a T cell epitope, might be a general explanation for the role of TG2 in celiac disease and a possible mechanism for the generation of autoantigens.

• Authors: Hanne Skovbjerg, Claus Koch, Dorit Anthonsen, Hans Sjöström
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1690 Issue 3 Pg. 220-30 (Nov 05 2004) ISSN: 0006-3002 [Print] Netherlands
• PMID: 15511629 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Albumins
• Amides
• Peptide Fragments
• Serum Albumin, Bovine
• Gliadin
• Transglutaminases
• Lysine

Topics

• Albumins (isolation & purification, metabolism)
• Amides (chemistry, metabolism)
• Amino Acid Sequence
• Animals
• Catalysis
• Cattle
• Celiac Disease (enzymology, metabolism)
• Deamination
• Gliadin (chemistry, metabolism)
• Guinea Pigs
• Intestinal Mucosa (chemistry, enzymology, metabolism)
• Lysine (metabolism)
• Molecular Sequence Data
• Peptide Fragments (chemistry, metabolism)
• Serum Albumin, Bovine (isolation & purification, metabolism)
• Spectrum Analysis
• Swine
• Transglutaminases (isolation & purification, metabolism)