Abstract
Protein kinase C from the anoxia-tolerant turtle Pseudemys scripta elegans was investigated to determine its role in mediating changes in brain metabolism associated with anoxia. Measurements of protein kinase C distribution in cytosol and membrane-associated fractions of cerebrum and hindbrain were performed with warm (18 °C)- and cold (7 °C)-acclimated animals exposed to normoxic or anoxic conditions. In cerebrum, the percentage of bound protein kinase C decreased from 48.5% to 35.1% in warm-acclimated animals and from 45.0% to 25.6% in cold-acclimated animals. In the hind-brain, bound protein kinase C increased from 45.0% to 72.9% in warm-acclimated animals and from 40.3% to 68.8% in cold-acclimated animals. The presence of three distinct protein kinase C isozymes (Types I, II and III) was confirmed by hydroxylapatite chromatography. The distribution of isozymes between cytosolic and membrane-associated fractions in cerebrum was 24% I, 37% II and 39% III (cytosolic) and 32% I, 35% II and 34% III (membrane-associated). In the hindbrain, the protein kinase C isozyme distribution was 34% I, 40% II and 26% III (cytosolic) and 18% I, 47% II and 35% III (membrane-associated). Kinetic characterization of the three isozymes showed that Type I was 27% activated by Ca2+, whereas Types II and III were only 4% and 2% activated by Ca2+, respectively. Full activity for all enzymes was observed only in the presence of phosphatidylserine and diacylglycerol. No differences in the K m for ATP, the K a for Ca2+ or the K a for phosphatidylserine were observed.
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Abbreviations
- EDTA:
-
ethylenediaminetetra-acetic acid
- EGTA:
-
ethyleneglycol-bis-(2-aminoethyl)-tetraacetic acid
- H7:
-
1-(5-isoquinolinyl sulphonyl)-2-methyl-piperazine
- OAG:
-
1-oleoyl-2-acetyl-rac-glycerol
- PIPES:
-
piperazine-n,n′-bis(2-ethanesulfonic acid)
- PKC:
-
protein kinase C
- PS:
-
phosphatidylserine
- TCA:
-
trichloroacetic acid
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Brooks, S.P.J., Storey, K.B. Protein kinase C in turtle brain: changes in enzyme activity during anoxia. J Comp Physiol B 163, 84–88 (1993). https://doi.org/10.1007/BF00309670
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DOI: https://doi.org/10.1007/BF00309670