Transamidating enzymes

Coagulation Factor XIIIa Undergoes a Conformational Change Evoked by Glutamine Substrate

“…Assays of XIIIa Activities Using Dimethylcasein As the Glutamine Substrate and Dansylcadaverine As the Amine Substrate-These assays followed continuous changes in fluorescence arising from the incorporation of dansylcadaverine into casein by the method of Lorand et al (27). The fluorescence was measured in 96-well microplates using a Perkin-Elmer luminescence spectrometer LS50, with excitation elicited at ex ϭ 360 nm and emission followed at em ϭ 510 nm.…”

Effects of Ascorbic Acid on the Formation Process for a Heat-induced Gel of Fish Meat (Kamaboko)

“…TGase activity was determined in terms of the incorporation of monodansylcadaverine (MDC) into N-Z-Amine AS (casein enzymatic hydrolysate from Sigma Chemical Co.) by a modified procedure of Lorand et al 29 ) The reaction mixture contained 2 mg/ml of N-ZAmine AS, 100,l1M MDC, 5 mM CaCI 2 , 0 or 10 mM dithiothreitol (DTT), o or 0.2% AsA, an appropriate amount of crude TGase (0.7-1.5 mg of protein), and 50mM Tris-HCI (pH 7.0) in a total volume of 3m!. The increase of fluorescence during incubation at 37°C was measured with a Hitachi 650-10S fiuoresence spectrophotometer at an excitation of 350 nm and an emission of 480 nm.…”

Unique Substrate Specificities of Two Adjacent Glutamine Residues in EAQQIVM for Transglutaminase: Identification and Characterization of the Reaction Products by Electrospray Ionization Tandem Mass Spectrometry

“…The amine-donor substrate selected for the FibN modification was monodansylcadaverine (MDC). MDC is a useful fluorescent probe for studying enzyme kinetics of TGase (17,18). To isolate and characterize the reaction products between FibN and MDC, reversed-phase HPLC (RP-HPLC) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) were carried out.…”