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Fig. 5 | Translational Neurodegeneration

Fig. 5

From: Serial deletion reveals structural basis and stability for the core enzyme activity of human glutaminase 1 isoforms: relevance to excitotoxic neurodegeneration

Fig. 5

Dual N- and C-terminal deletion of GAC and KGA. a) Schematic illustration of the dual N- and C-terminal deletion on GAC and KGA was shown. b) Glutaminase activity of each of the mutant was determined by the enzyme activity assay with varying glutamine concentrations. Data was plotted as mmol/L per 30 min reaction time against the glutamine concentration. Non-linear regression was performed with GraphPad Prism software. Results shown are the means ± SD of triplicate samples. c) Full-length GAC/KGA, as well as the truncated 219–550 AA GAC/KGA protein were expressed in BL21 cells. The cells were cultured to reach optimal OD values (0.6–0.7) before protein production was induced by IPTG. Glutamate levels in the extracellular fluid were determined by RP-HPLC. Protein purification of the crude lysate derived from the empty vector transfected BL21 cells served as the control. Results shown are the means ± SD of triplicate samples. FL, full-length; NS, non-significant; *, p < 0.05, compared with the activities of FL GAC and KGA

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