title: Proteomics identification of acyl-acceptor and acyl-donor substrates for transglutaminase in a human intestinal epithelial cell line. Implications for celiac disease.
abstract: Transglutaminase (TG)-catalyzed cross-linking of both intracellular and extracellular proteins is an important biochemical event. However, increased concentrations of cross-linked proteins have been observed in many disorders. Moreover, TG-catalyzed modification of proteins might generate new self-antigens responsible for the autoimmune response, as in celiac disease. The identification of available substrates may offer an understanding of how the TG-catalyzed post-translational modification has an impact on physiology and disease. We used a proteomic approach to identify TG-modified protein targets in human intestinal epithelial cells to determine the extent to which transglutaminase specifically contributes to celiac disease. Two probes were used for endogenous TG activity: 5-(biotinamido)pentylamine, which represents the acyl-acceptor, and a biotinylated glutamine-containing peptide, which represents the acyl-donor. This approach identified >25 proteins, which range from 30,000 to 300,000 Daltons and can serve as acyl-acceptor and/or acyl-donor for transglutaminase. Some of them were known transglutaminase substrates, whereas others had not been previously identified. These targets include proteins involved in cytoskeletal network organization, folding of proteins, transport processes, and miscellaneous metabolic functions.
authors.affiliation: Department of Chemistry, University of Salerno, 84081 Baronissi, Salerno, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Orrù
authors.forename: Ivana
authors.initials: I
authors.lastname: Caputo
authors.forename: Alfonsina
authors.initials: A
authors.lastname: D'Amato
authors.forename: Margherita
authors.initials: M
authors.lastname: Ruoppolo
authors.forename: Carla
authors.initials: C
authors.lastname: Esposito
mesh_terms: D000818:Animals; D002446:Celiac Disease / Q000201:enzymology / Q000378:metabolism*; D002460:Cell Line; D004285:Dogs; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D040901:Proteomics; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D011503:Transglutaminases