Phosphorylation of the α-amino-3-hydroxy-5-methylisoxazole-4-propionic Acid receptor GluR1 subunit by calcium/calmodulin-dependent kinase II

Andrew L. Mammen, Kimihiko Kameyama, Katherine W. Roche, Richard L Huganir

Research output: Contribution to journalArticle

Abstract

Modulation of α-amino-3-hydroxy-5-methylisoxazole-4-propionic Acid (AMPA) receptors in the brain by protein phosphorylation may play a crucial role in the regulation of synaptic plasticity. Previous studies have demonstrated that calmodulin (CAM) kinase II can phosphorylate and modulate AMPA receptors. However, the sites of CaM kinase phosphorylation have not been unequivocally identified. In the current study, we have generated two phosphorylation site-specific antibodies to analyze the phosphorylation of the glutamate receptor GluR1 subunit. These antibodies recognize GluR1 only when it is phosphorylated on serine residues 831 or 845. We have used these antibodies to demonstrate that serine 831 is specifically phosphorylated by CaM kinase II in transfected cells expressing GluR1 as well as in hippocampal slice preparations. Two-dimensional phosphopeptide mapping experiments indicate that Set-831 is the major site of CaM kinase II phosphorylation on GluR1. In addition, treatment of hippocampal slice preparations with phorbol esters and forskolin increase the phosphorylation of serine 831 and 845, respectively, indicating that protein kinase C and protein kinase A phosphorylate these residues in hippocampal slices. These results identify the site of CaM kinase phosphorylation of the GluR1 subunit and demonstrate that GluR1 is multiply phosphorylated by protein kinase A, protein kinase C, and CaM kinase II in situ.

Original languageEnglish (US)
Pages (from-to)32528-32533
Number of pages6
JournalJournal of Biological Chemistry
Volume272
Issue number51
DOIs
StatePublished - Dec 19 1997

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Calcium-Calmodulin-Dependent Protein Kinases
Phosphorylation
Calmodulin
Phosphotransferases
Calcium
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Serine
Cyclic AMP-Dependent Protein Kinases
Antibodies
Phosphopeptides
Neuronal Plasticity
Glutamate Receptors
Phorbol Esters
Colforsin
propionic acid
bucide
Protein Kinase C
Plasticity
Brain
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of the α-amino-3-hydroxy-5-methylisoxazole-4-propionic Acid receptor GluR1 subunit by calcium/calmodulin-dependent kinase II. / Mammen, Andrew L.; Kameyama, Kimihiko; Roche, Katherine W.; Huganir, Richard L.

In: Journal of Biological Chemistry, Vol. 272, No. 51, 19.12.1997, p. 32528-32533.

Research output: Contribution to journalArticle

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