Contribution of Special Structural Features to High Thermal Stability of a Cold-Active Transglutaminase.

A cold-active transglutaminase (TGase, EC 2.3.2.13) that catalyzes the reaction of protein glutamine + protein lysine ↔ protein with γ-glutamyl-ε-lysine cross-link + NH3 at low temperatures was reported previously. This study verified the thermal stability of the TGase from 0-80 °C. Fluorescence and CD spectra studies confirmed tertiary structural damage at 40 °C, α-helix reduction at 60 °C, and refolding during cooling to 20 °C. The TGase sequence was obtained by transcriptomics and used to build its structure. Its catalytic triad was Cys333-His403-Asp426 and its catalytic process was inferred from the model. Molecular dynamics simulation illustrated that its cold activity resulted from its flexible active site, while high thermostability was conferred by an overall rigid structure, a large amount of stable Val and Lys, and strong electrostatic interactions at the N- and C- terminals. This study fills gaps in the correlation of conformational changes with stability and activity of TGase.