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Molecular Characterization of Covalent Complexes between Tissue Transglutaminase and Gliadin Peptides

Authors :
Peter Roepstorff
Ludvig M. Sollid
Burkhard Fleckenstein
Günther Jung
Martin R. Larsen
Shuo-Wang Qiao
Source :
Fleckenstein, B, Qiao, S-W, Larsen, M R, Jung, G, Roepstorff, P & Sollid, L M 2004, ' Molecular characterization of covalent complexes between tissue transglutaminase and gliadin peptides ', Journal of Biological Chemistry, vol. 279, no. 17, pp. 17607-16 . https://doi.org/10.1074/jbc.M310198200
Publication Year :
2004
Publisher :
Elsevier BV, 2004.

Abstract

Tissue transglutaminase (TG2) modifies proteins and peptides by transamidation or deamidation of specific glutamine residues. TG2 also has a central role in the pathogenesis of celiac disease. The enzyme is both the target of disease-specific autoantibodies and generates deamidated gliadin peptides recognized by intestinal T cells from patients. Incubation of TG2 with gliadin peptides also results in the formation of covalent TG2-peptide complexes. Here we report the characterization of complexes between TG2 and two immunodominant gliadin peptides. Two types of covalent complexes were found; the peptides are either linked via a thioester bond to the active site cysteine of TG2 or via isopeptide bonds to particular lysine residues of the enzyme. We quantified the number of gliadin peptides bound to TG2 under different conditions. After 30 min of incubation of TG2 at 1 microm with an equimolar ratio of peptides to TG2, approximately equal amounts of peptides were bound by thioester and isopeptide linkage. At higher peptide to TG2 ratios, more than one peptide was linked to TG2, and isopeptide bond formation dominated. The lysine residues in TG2 that act as acyl acceptors were identified by matrix assisted laser desorption ionization and nanoelectrospray mass spectrometry and tandem mass spectrometry analysis of proteolytic digests of the TG2-peptide complexes. At a high molar excess of gliadin peptides to TG2 altogether six lysine residues of TG2 were found to participate in isopeptide bond formation. The results are relevant to the understanding of how antibodies to TG2 are formed in celiac disease.

Details

ISSN :
00219258
Volume :
279
Database :
OpenAIRE
Journal :
Journal of Biological Chemistry
Accession number :
edsair.doi.dedup.....38912079b03a483f6b0f5b9dd4f76cdc
Full Text :
https://doi.org/10.1074/jbc.m310198200