GGCT is a 188 amino acid protein that consists of six beta-strands, five alpha-helices, and four short 3-10 helices . The enzyme’s activity is measured by its ability to release L-alanine from gamma-glutamyl-L-alanine, with the detection of alanine by alanine dehydrogenase . The specific activity of GGCT is greater than 40,000 pmol/min/μg under the described conditions .
GGCT plays a significant role in maintaining glutathione homeostasis, which is vital for cellular defense against oxidative stress and detoxification of xenobiotics . The enzyme’s function is critical in various physiological processes, including the regulation of apoptosis and cellular proliferation .
Up-regulation of GGCT expression has been observed in various tumor tissues, including lung, esophagus, stomach, bile duct, uterine cervix, colon, and breast . This makes GGCT a potential biomarker for numerous types of cancers. The enzyme’s involvement in the gamma-glutamyl cycle and its role in apoptosis highlight its importance in cancer biology and potential therapeutic targeting .
Recombinant human GGCT is produced using E. coli as the expression system . The recombinant enzyme is purified to greater than 95% purity by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain . The endotoxin level is less than 1.000 EU per 1 µg of the protein by the LAL method . The recombinant enzyme is supplied as a 0.2 μm filtered solution in Tris and NaCl and is stable when stored at -70°C .