Affinity purification, identification, and biochemical characterization of Gamma-glutamyl transpeptidase, a membrane anchored enzyme of Gigantocotyle explanatum

Abstract

Gamma-glutamyl Transpeptidase is an enzyme that facilitates the transfer of glutamyl groups from glutamyl peptides to other peptides or water. Additionally, it also participates in important processes such amino acid transport, cellular redox control, drug detoxi cation, apoptosis, and DNA fragmentation in a various organism. In the present study, GGT activity in Gigantocotyle explanatum was examined in order to characterize the enzyme in the helminth system. GGT is isolated using membrane solubilization and puri ed through a nity column chromatography (Con-A Sepharose column). Km and Vmax values, as well as the optimal pH, optimal temperature, and incubation period, are also determined using enzyme kinetics. The hetero-dimeric property of the enzyme is demonstrated by the puri ed GGT, which yielded two subunits of 65.5 and 55 kDa. The optimal pH and temperature are found to be 8.0 and 37°C, respectively. While assessing the optimal incubation time of the enzyme, it was observed that the puri ed GGT not only retained its functional integrity up to 15 min but also re ected considerable thermostability at higher temperatures, by retaining 78% and 25% of its initial activities at 50°C and 60°C, respectively. 1 mM concentration of 6-Diazo-5-Oxo Nor-isoleucine (DON), a speci c inhibitor of GGT, completely abolished GGT activity. These results suggest that GGT in these worms is a catalytically active enzyme with distinguishing characteristics that can be used for further study to comprehend its function in amphistome biology and in host-parasite relationships, especially since the potential therapeutic candidacy of the GGT enzyme has already been indicated in these groups of organisms.

The Augural Interplay of Myosteatosis and Gamma-Glutamyltransferase in Patients Undergoing Comprehensive Surgical Intervention for Cholangiocarcinoma

“…To this juncture, explorations delving into the underpinnings of the correlation between gamma-glutamyl transferase (GGT) and adverse prognoses in malignant neoplasms remain regrettably sparse. Esteemed as a mercaptan-rich enzyme, GGT is most profuse within the renal system, trailed by the pancreas, pulmonary organs, and hepatic structures, where it partakes in the conveyance of glutamine molecules throughout the organism [29,30]. The current corpus of scienti c literature posits that GGT plays a cardinal role in contributing to unfavorable outcomes in oncology patients by stimulating the generation of reactive oxygen species (ROS) within the neoplastic microcosm, thereby spurring the proliferation, expansion, and in ltration of the malignant growth [31].…”