Molecular basis of protein cross-linking reactions for cellular functions

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Transglutaminases are the enzymes that catalyze protein cross-linking or modification via peptide-bound glutamine residues. These reactions are essential for the multiple biological processes such as blood coagulation, skin epidermis formation and apoptosis. In order to elucidate the physiological roles and expression sites on mammals, using highly reactive substrate peptides (obtained by us for these years), we analyzed expression pattern and established identification system of possible substrates. In the studies, expressions for RNA, protein (using antibody), in situ enzymatic activity were investigated. Furthermore, establishment of FRET system to detect intracellular activity was attempted. As a result, in vitro system was successful. In addition, Medaka (Oryzias latepis) as a model animal, was analyzed for the orthologues of transglutaminases: biochemical data and establishment of a couple of genetic loss fish was produced.