Thermal Stability of Glucokinases in Thermoanaerobacter tengcongensis
Figure 3
In vitro (a) and in vivo (b) protein degradation assays of ATP- and ADP-GLK by Western blot. (a) Recombinant ATP- or ADP-GLK was incubated with (T) or without (C) T. tengcongensis native protein lysate at 75°C or 80°C for 60 min. The reaction was then stopped by addition of SDS loading buffer. After SDS-PAGE separation, the remaining protein was detected by Western blot with specific anti-His6 antibody. (b) T. tengcongensis cells cultured at 75°C to log phase (set as time 0 h) were transferred to 80°C for further culturing and then collected at the indicated time points to get soluble proteins for Western blot analysis with specific anti-ATP-GLK or anti-ADP-GLK antibodies.
