Depolarisation-dependent protein phosphorylation in rat cortical synaptosomes: factors determining the magnitude of the response.
The sequence of molecular events linking depolarisation-dependent calcium influx to the release of neurotransmitters from nerve terminals is unknown; however, calcium-stimulated protein phosphorylation may play a role. In this study the incorporation of phosphate into proteins was investigated using an intact postmitochondrial pellet isolated from rat cerebral cortex. The rate and relative incorporation of label into individual phosphoproteins depended on the prelabelling time and buffer concentrations of calcium and phosphate. After prelabelling for 45 min, depolarisation caused a greater than 20% increase in the labelling of 10 phosphoproteins, and this initial increase was maximal with 41 mM K+ for 5 s, or 30 microM veratridine for 15 s, in the presence of 1 mM calcium. Both agents also led to an initial dephosphorylation of four phosphoproteins. Depolarisation for 5 min led to a significant decrease in the labelling of all phosphoproteins. All of the depolarisation-stimulated changes in protein phosphorylation were calcium-dependent. The depolarisation conditions found to optimally alter the phosphorylation of synaptosomal proteins find many parallels in studies on calcium uptake and neurotransmitter release. However, the uniform responses of such a large number of phosphoproteins to the multitude of depolarisation conditions studied suggest that the changes could equally well relate to recovery events such as biosynthesis of neurotransmitters and regulation of intraterminal metabolic activity.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002540:Cerebral Cortex / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D007700:Kinetics; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013574:Synaptosomes / Q000502:physiology*; D014701:Veratridine / Q000494:pharmacology
D010750:Phosphoproteins; D014701:Veratridine; D011188:Potassium; D002118:Calcium