Phosphorylation of synaptic membranes

Robert F Berman, J. P. Hullihan, W. J. Kinnier, J. E. Wilson

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19 Scopus citations

Abstract

In vivo and in vitro phosphorylation of synaptic membrane proteins were compared. In vivo phosphorylation was carried out by injecting rats intraventricularly with 1 mCi of 32P1-orthophosphate. After a 40-min isotope incorporation period, rats were decapitated and the synaptic membranes isolated. In vitro phosphorylation was accomplished by incubating unlabeled synaptic membranes, isolated from noninjected rats, with 5 μM-[γ-32P]ATP. In vivo and in vitro 32 P-labeled synaptic membranes were then fractionated electrophoretically on an SDS-polyacrylamide (7.5%) slab gel. The Coomassie blue protein patterns for in vivo and in vitro 32 P-labeled synaptic membranes were identical. In contrast, autoradiographs of these gels showed striking differences in the pattern of phosphate incorporation. Cyclic-AMP (10 μM) maximally stimulated the in vitro phosphate labelling of two protein bands (80,000 and 55,000 apparent molecular weight) which were not substantially labeled by the in vivo procedure.

Original languageEnglish (US)
Pages (from-to)431-437
Number of pages7
JournalJournal of Neurochemistry
Volume34
Issue number2
StatePublished - 1980
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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    Berman, R. F., Hullihan, J. P., Kinnier, W. J., & Wilson, J. E. (1980). Phosphorylation of synaptic membranes. Journal of Neurochemistry, 34(2), 431-437.