Purification of transglutaminase from scallop [Patinopecten yessoensis] striated adductor muscle and NaCl-induced inactivation

Tissue type transglutaminase (TGase) was purified from scallop striated adductor muscle with successive chromatographies of DE-52 cellulose, Sephacryl S-300, and Mono Q columns. The yield and purification of the enzymatic activity was 16.6% and 101.9-fold, respectively. The molecular mass of purified enzyme was estimated to be 95 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. Scallop TGase was Ca sup(2+)-dependent and strongly inactivated by p-chloromercuribenzoic acid, N-ethylmaleimide, Cu sup(2+), and Zn sup(2+), meaning it belongs to the thiol group of enzymes as well as being a mammalian enzyme. When scallop TGase was incubated in 0.5 M NaCl without substrate for 2 h at 20 deg C and pH7.5, enzymatic activity decreased to 14.4% of its original. However, a conformational change in the TGase molecule was not detected by either fluorescent, ultraviolet, and circular dichroism spectra analyses compared to the enzyme incubated without NaCl. In addition, the enzyme inactivated by NaCl was partially recovered by the dilution of salt concentration, which means that the NaCl-induced inactivation process is reversible to some extent. These results suggest that NaCl-induced modulation of the TGase molecule occurs via a small conformational change.