A role for tissue transglutaminase in alpha-gliadin peptide cytotoxicity.
In coeliac disease, gliadin peptides p56-88, p57-68 and p31-49 have been demonstrated to be involved in the pathogenic damage of the small intestine via their immunogenicity or toxicity to epithelial cells. To try to understand the mechanism of their toxicity, we investigated the effect of synthetic peptides (p31-49, p56-88, p57-68, p69-82) and of their deamidated analogues on Caco2 and FHs 74 Int cell toxicity and tissue tranglutaminase activity. Apoptosis, necrosis and cell viability were assessed by flow cytometry, and peptide deamidation was determined indirectly by measuring its capacity to inhibit tTG activity. The results showed that p56-88 and p57-68 reduced cell growth and concomitantly inhibited tTG activity in both cell types. This effect was abolished when Caco2 cells were treated with antibodies to tTG. Deamidated peptide p57-68 (E(65)) lost practically all of its inhibitory effect on cell growth and on tTG activity. Cellular toxicity was also observed with p31-49, which was not a substrate for tTG. p69-82 was not cytotoxic but became so when glutamine 72 was substituted by glutamic acid. These findings provide evidence for the existence of three types of toxicity among gliadin peptides: (i) peptides that are intrinsically toxic and are not substrates of tTG; (ii) peptides that are non-toxic but become so when they act as substrates of tTG; and (iii) peptides that are non-toxic and are not substrates of tTG but become so when deamidated. A mechanism other than that involving tTG could be responsible for the deamidation of glutamine residues of gliadin in the intestinal tract.
Sakly
W
W
Covalab, Villeurbanne, France.
Thomas
V
V
Quash
G
G
El Alaoui
S
S
D018938:Caco-2 Cells; D002446:Celiac Disease / Q000201:enzymology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003641:Deamination; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005903:Gliadin / Q000494:pharmacology*; D006801:Humans; D010446:Peptide Fragments / Q000494:pharmacology; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*
D004791:Enzyme Inhibitors; D010446:Peptide Fragments; D005903:Gliadin; D011503:Transglutaminases