TY - JOUR
T1 - Contribution of Special Structural Features to High Thermal Stability of a Cold-Active Transglutaminase
AU - Zhang, Yi
AU - Li, Chen
AU - Geary, Timothy
AU - Simpson, Benjamin Kofi
N1 - Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/7/29
Y1 - 2020/7/29
N2 - 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.
AB - 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.
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U2 - 10.1021/acs.jafc.0c03344
DO - 10.1021/acs.jafc.0c03344
M3 - Article
C2 - 32643372
AN - SCOPUS:85089610547
SN - 0021-8561
VL - 68
SP - 7935
EP - 7945
JO - Journal of agricultural and food chemistry
JF - Journal of agricultural and food chemistry
IS - 30
ER -