title: Transglutaminase 2 and Factor XIII catalyze distinct substrates in differentiating osteoblastic cell line: utility of highly reactive substrate peptides.
abstract: Differentiated osteoblastic cell line, MC3T3-E1 expresses transglutaminase 2 (TG2) and Factor XIII (FXIII). In previous studies, we identified isozyme-specific and highly reactive glutamine-donor substrate peptides (pepF11KA and pepT26) for each isozyme. Using these peptides, we compared the reaction products with lysine-donor substrates for each isozyme in differentiating MC3T3-E1 cells. By this analysis, distinct substrates for the activated TG2 and FXIII were detected in cultured cellular extract. Possible substrates that incorporated biotin-labeled peptides were further purified using streptavidin-affinity chromatography. Several isozyme-specific substrates were identified by mass spectrometry analysis of the purified fractions. These analyses also indicate the benefit of the substrate peptides for obtaining distinct substrates in a reaction mixture where two isozymes co-exist.
authors: Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 4648601, Japan.
authors: Kazuya
authors: K
authors: Watanabe
authors: Kanako
authors: K
authors: Tsunoda
authors: Miho
authors: M
authors: Itoh
authors: Mina
authors: M
authors: Fukui
authors: Hitoshi
authors: H
authors: Mori
authors: Kiyotaka
authors: K
authors: Hitomi
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002454:Cell Differentiation; D005176:Factor XIII / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D051379:Mice; D010006:Osteoblasts / Q000201:enzymology* / Q000502:physiology; D010446:Peptide Fragments / Q000378:metabolism; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D025941:Protein Interaction Mapping; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D010446:Peptide Fragments; D005176:Factor XIII; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins