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Analysis of transglutaminase protein substrates by functional proteomics
Author(s) -
Ruoppolo Margherita,
Orrù Stefania,
D'Amato Alfonsina,
Francese Simona,
Rovero Paolo,
Marino Gennaro,
Esposito Carla
Publication year - 2003
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.0239103
Subject(s) - tissue transglutaminase , biotinylation , lysine , biochemistry , proteomics , enzyme , chemistry , peptide , amino acid , biotin , gene
Abstract Transglutaminases are calcium‐dependent enzymes that catalyze a post‐translational modification of proteins through the formation of ε‐(γ‐glutamyl)lysine bonds. Although specific roles for transglutaminases have been described, recent findings have provided evidence that dysregulation of transglutaminases may contribute to many pathological processes including celiac disease and neurodegenerative diseases. A crucial step in the elucidation of biological and pathological roles of transglutaminases requires the identification of protein substrates. A strategy based on a functional proteomic analysis was set up using two well‐characterized biotinylated transglutaminase substrates as affinity probes: 5–(biotinamido)pentylamine and the synthetic biotinylated peptide TVQQEL, the amino‐ and acyl‐donor probes, respectively. A pool of known tissue type transglutaminase protein substrates was selected in order to test the procedure. Results obtained in this paper indicate that the whole strategy can be successfully applied in order to identify transglutaminases protein substrates as well as the amino acid site sensitive toward enzyme activity.