The effect of hibernation on protein phosphatases from ground squirrel organs

Protein phosphorylation has been identified as a reversible mechanism for the regulated suppression of metabolism and thermogenesis during mammalian hibernation. The effects of hibernation on the activity of serine/threonine and tyrosine protein phosphatases (PP1, PP2A, PP2C and PTPs) were assessed in five organs of Richardson's ground squirrel. Each phosphatase subfamily responded differently during torpor, and each showed organ-specific patterns of activity changes. The distribution of PP1 catalytic subunit (PP1c) isoforms (alpha, delta, gamma1) was assessed in five organs, and changes in the subcellular distribution of PP1 were observed during hibernation in liver and muscle. For example, in muscle, cytosolic PP1 content increased and myofibril-associated PP1 decreased during torpor. PP1c from ground squirrel liver was purified to homogeneity and characterized; temperature effects on PP1c maximal activity suggested that temperature had little or no effect on relative dephosphorylation potential at low temperatures. However, nucleotide inhibition of PP1c by ATP, ADP and AMP was much weaker at 5 degrees C compared with 37 degrees C assay temperatures. PP2A activity decreased in three organs (brown adipose, kidney, brain) during hibernation whereas PP2C activity was increased in liver and brain. PTPs were assessed using both a general substrate (ENDpYINASL) and a substrate (DADEpYLIPQQG) specific for PTPs containing the SH2-binding site; both revealed hibernation-associated changes in PTP activities. Changes in protein phosphatase activities suggest the relative importance of these modules in controlling metabolic function and cellular processes during mammalian hibernation.