Abstract
We have investigated the possible role of plasma membrane oxidoreductases in the Ca2+ export mechanisms in rat brain synaptic membranes. Ca2+ efflux in nerve terminals is controlled both by a high-affinity/low capacity Mg-dependent ATP-stimulated Ca2+ pump and by a low affinity/high capacity ATP-independent Na+-Ca2+ exchanger. Both Ca2+ efflux mechanisms were strongly inhibited by pyridine nucleotides, in the order NADP>NAD>NADPH>NADH with IC50 values of ca. 10 mM for NADP and ca. 3 mM for the other agents in the case of the ATP-driven Ca2+ pump and with IC50 values between 8 and 10 mM for the Na+-Ca2+ exchanger. Oxidizing agents such as DCIP and ferricyanide inhibited the ATP-driven Ca2+ efflux mechanism but not the Na+-Ca2+ exchanger. In addition, full activation of plasma membrane oxidoreductases requires both an acceptor and an electron donor; therefore the combined effects of both substrates added together were also studied. When plasma membrane oxidoreductases of the synaptic plasma membrane were activated in the presence of both NADH (or NADPH) and DCIP or ferricyanide, the inhibition of the ATP-driven Ca2+ pump was optimal; by contrast, the pyridine nucleotide-mediated inhibition of the Na+-Ca2+ exchanger was partially released when both substrates of the plasma membrane oxidoreductases were present together. Furthermore, the activation of plasma membrane oxidoreductases also strongly inhibited intracellular protein phosphorylation in intact synaptosomes, mediated by eithercAMP-dependent protein kinase, Ca2+ calmodulin-dependent protein kinases, or protein kinase C.
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Abbreviations
- Hepes:
-
4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid
- SDS:
-
sodium dodecyl sulfate
- EGTA:
-
ethylenglycol-bis(β-aminoethylether)-N,N,N′,N′-tetraacetic acid
- DCIP:
-
dichlorophenol-indophenol
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Bulliard, C., Marmy, N. & Dreyer, J.L. Effects of plasma membrane oxidoreductases on Ca2+ mobilization and protein phosphorylation in rat brain synaptosomes. J Bioenerg Biomembr 22, 645–662 (1990). https://doi.org/10.1007/BF00809069
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DOI: https://doi.org/10.1007/BF00809069