In vivo protein phosphorylation was examined in postsynaptic density-enriched fractions isolated from rat brain. In vivo phosphorylation was carried out by injecting rats intraventricularly with [32P]orthophosphate followed by isolation of postsynaptic densities from pooled cerebral cortices. In vivo 32P-labeled postsynaptic densities were then fractionated by sodium dodecylsulfate-polyacrylamide slab gel electrophoresis and stained with Coomassie Blue.
View Article and Find Full Text PDF1--The longitudinal muscle-myenteric plexus strip preparation of the guinea-pig ileum was used to study the binding of [3H]-diazepam and the effect of benzodiazepines on its contraction. 2--Scatchard analysis of binding indicated a single class of binding sites with KD = 43 nM and Bmax = 229 fmol/mg prótein. Binding was of peripheral type based on the much greater binding affinity of Ro5-4864 as compared to clonazepam.
View Article and Find Full Text PDFA 3200-dalton adrenal enkephalin-containing peptide, designated peptide E, that exhibits high opiate activity in the guinea pig ileum longitudinal muscle preparation was purified, and its structure was determined. It contains an amino-terminal [Met]enkephalin sequence and a [Leu]enkephalin sequence at the carboxyl terminus. Sequence analysis revealed that peptide E arises from a previously characterized 4900-dalton adrenal enkephalin-containing peptide.
View Article and Find Full Text PDFDopamine (3-hydroxytyramine) stimulates the incorporation of 32P into proteins endogenous to a homogenate of rat caudate nucleus when 10 microM [gamma-32P]-ATP is used as a substrate following preincubation with 400 microM ATP. The increase in 32P incorporation has pharmacological characteristics similar to caudate tissue. Chronic depletion of striatal dopamine in vivo by stereotaxic injection of 6-hydroxydopamine in the nigrostriatal pathway results in a significant enhancement of the dopamine stimulation of 32 p incorporation in vitro.
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