Skeletal muscle hexokinase: regulation in mammalian hibernation.
Skeletal muscle hexokinase (HK) from Richardson's ground squirrels was analyzed to determine how the enzyme is regulated during hibernation, a state of cold torpor. The HK II isozyme dominated in muscle and ~15% of total HK was bound to the insoluble fraction. HK maximum activity was 33% lower in hibernator muscle and the enzyme showed a significantly higher K ( m ) ATP (by 80%) and a lower K ( i ) for glucose-6-P (by 40%) than euthermic HK (assayed at 22 degrees C). However, 5 degrees C assay significantly reduced K ( m ) glucose of hibernator HK. Stimulation of AMP-dependent protein kinase (AMPK) in hibernator extracts elevated the HK activity and reduced K ( m ) ATP, but did not affect euthermic HK. Stimulation of protein phosphatases significantly lowered the HK activity in both situations. AMPK-dependent phosphorylation was confirmed by immunopreciptiation of (32)P-labeled HK. DEAE-Sephadex ion exchange chromatography revealed two peaks of HK in hibernator muscle extracts (low and high phosphate forms), whereas only a single peak of phospho-HK was present in euthermic muscle. We conclude that differential control of muscle HK in euthermic versus hibernating states is derived from two main regulatory influences, reversible protein phosphorylation and temperature effects on kinetic properties.
Abnous
Khalil
K
Institute of Biochemistry and Department of Chemistry, Carleton University, Ottawa, ON, Canada.
Storey
Kenneth B
KB
D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D019298:Glucose-6-Phosphate / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism*; D006605:Hibernation / Q000502:physiology*; D018482:Muscle, Skeletal / Q000201:enzymology*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation / Q000502:physiology; D012377:Rodentia / Q000378:metabolism*
D019298:Glucose-6-Phosphate; D000255:Adenosine Triphosphate; D006593:Hexokinase; D055372:AMP-Activated Protein Kinases; D010749:Phosphoprotein Phosphatases