Cyclic AMP-dependent protein kinase does not increase calcium transport in platelet microsomes.
Cyclic AMP inhibits platelet activation, at least in part, by reducing intracellular levels of ionic calcium. Previous studies using platelet microsomal fractions have suggested that one mechanism for this effect is stimulation by cyclic AMP and its protein kinase of calcium uptake into microsomal storage sites. In the present study, the effect of cyclic AMP and its protein kinase on calcium uptake by microsomal membranes has been re-examined using the active catalytic subunit of cyclic AMP-dependent protein kinase. The catalytic subunit increased calcium uptake two-fold, but this effect was not inhibited by boiling the catalytic subunit or by recombination with the regulatory subunit of cyclic AMP-dependent protein kinase, conditions that inhibited catalytic subunit activity. Conversely, dialysis of the catalytic subunit preparation against low phosphate buffer, which did not inhibit catalytic subunit activity, inhibited the stimulation of calcium uptake by the catalytic subunit preparation. Finally, the addition of high phosphate buffer, similar in phosphate concentration to that of the catalytic subunit preparation, stimulated calcium uptake. We conclude that the catalytic subunit does not directly stimulate calcium uptake by platelet microsomes.
White
G C
GC
Department of Medicine, University of North Carolina, Chapel Hill 27514.
Barton
D W
DW
White
T E
TE
Fischer
T H
TH
D001692:Biological Transport / Q000502:physiology; D001792:Blood Platelets / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D002118:Calcium / Q000097:blood*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D066298:In Vitro Techniques; D008861:Microsomes / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000502:physiology*
D001798:Blood Proteins; D011494:Protein Kinases; D002118:Calcium