Glutamate dehydrogenase (GDH) was purified to homogeneity from the liver of euthermic (37°C body temperature) and hibernating (torpid, 5°C body temperature) Richardson's ground squirrels (Spermophilus richardsonii). SDS-PAGE yielded a subunit molecular weight of 59.5 ± 2 kDa for both enzymes, but reverse phase and size exclusion HPLC showed native molecular weights of 335 ± 5 kDa for euthermic and 320 ± 5 kDa for hibernator GDH. Euthermic and hibernator GDH differed substantially in apparent Km values for glutamate, NH4 +, and α-ketoglutarate, as well as in Ka and IC50 values for nucleotide and ion activators and inhibitors. Kinetic properties of each enzyme were differentially affected by assay temperature (37 versus 5°C). For example, the Km for α-ketoglutarate of euthermic GDH was higher at 5°C (3.66 ± 0.34 mM) than at 37°C (0.10 ± 0.01 mM), whereas hibernator GDH had a higher affinity for α-ketoglutarate at 5°C (Km was 0.98 ± 0.08 mM at 37°C and 0.43 ± 0.02 mM at 5°C). Temperature effects on Ka ADP values of the enzymes followed a similar pattern; GTP inhibition was strongest with the euthermic enzyme at 37°C and weakest with hibernator GDH at 5°C. Entry into hibernation leads to stable changes in the properties of ground squirrel liver GDH that allow the enzyme to function optimally at the prevailing body temperature.

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Keywords Amino acid metabolism, Mammalian hibernation, Spermophilus richardsonii, Temperature-dependent enzyme kinetics
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Journal Biochemistry and Cell Biology
Thatcher, B.J., & Storey, K. (2001). Glutamate dehydrogenase from liver of euthermic and hibernating Richardson's ground squirrels: Evidence for two distinct enzyme forms. Biochemistry and Cell Biology, 79(1), 11–19. doi:10.1139/bcb-79-1-11