Purification and characterization of fructose bisphosphate aldolase from the ground squirrel, Spermophilus lateralis: enzyme role in mammalian hibernation.
Fructose-1,6-bisphosphate (F1,6P(2)) aldolase was purified to homogeneity from skeletal muscle of the golden-mantled ground squirrel, Spermophilus lateralis. Enzyme properties were examined at temperatures characteristic of euthermia (37 degrees C) and hibernation (5 degrees C); parallel studies assessed rabbit muscle aldolase for comparison. Kinetic properties of each enzyme were differentially affected by assay temperature. For example, the K(m) for F1,6P(2) of ground squirrel aldolase was 0.9+/-0.05 microM at 37 degrees C and 50% higher (1.45+/-0.04 microM) at 5 degrees C, whereas the K(m) of rabbit aldolase increased threefold over the same temperature range. The inhibitory effects of adenylates were similar at both temperatures for the ground squirrel enzyme, but inhibition by adenosine 5(')-diphosphate, adenosine 5(')-monophosphate, and inosine 5(')-monophosphate was substantially reduced at 5 degrees C for rabbit aldolase. Inhibition by inorganic phosphate increased at lower temperatures for both enzymes; for ground squirrel aldolase, the K(i) was 1.18+/-0.1mM at 37 degrees C and 0.23+/-0.05 mM at 5 degrees C. Inhibition of aldolase by inorganic phosphate could be one factor that helps to shut down glycolysis during hibernation. Thus, mammalian hibernators may exploit low-temperature characteristics of aldolase to benefit the metabolic needs of the hibernating state.
MacDonald
Justin A
JA
Institute of Biochemistry and Department of Biology, College of Natural Sciences, Carleton University, Ottawa, Ont., Canada.
Storey
Kenneth B
KB
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005634:Fructose-Bisphosphate Aldolase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D006605:Hibernation / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D007700:Kinetics; D018482:Muscle, Skeletal / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology; D011817:Rabbits; D012589:Sciuridae / Q000502:physiology*; D013696:Temperature
D004791:Enzyme Inhibitors; D010710:Phosphates; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D005634:Fructose-Bisphosphate Aldolase