Mapping the protein phosphatase-2B anchoring site on AKAP79. Binding and inhibition of phosphatase activity are mediated by residues 315-360

Mark L. Dell'Acqua, Kimberly L. Dodge, Steven Tavalin, John D. Scott

Research output: Contribution to journalArticle

116 Citations (Scopus)

Abstract

Compartmentalization of protein kinases and phosphatases with substrates is a means to increase the efficacy of signal transduction events. The A-kinase anchoring protein, AKAP79, is a multivalent anchoring protein that maintains the cAMP-dependent protein kinase, protein kinase C, and protein phosphatase-2B (PP2B/calcineurin) at the postsynaptic membrane of excitatory synapses where it is recruited into complexes with N-methyl-D-aspartic acid or α-amino-3-hydroxy-5-metbyl-isoxazole-4-propionic acid (AMPA)-subtype glutamate receptors. We have used cellular targeting of AKAP79 truncation and deletion mutants as an assay to map the PP2B-binding site on AKAP79. We demonstrate that residues 315-360 are necessary and sufficient for AKAP79-PP2B anchoring in cells. Multiple determinants contained within this region bind directly to the A subunit of PP2B and inhibit phosphatase activity. Peptides spanning the 315-360 region of AKAP79 can antagonize PP2B anchoring in vitro and targeting in transfected cells. Electrophysiological experiments further emphasize this point by demonstrating that a peptide encompassing residues 330-357 of AKAP79 attenuates PP2B-dependent down-regulation of GluR1 receptor currents when perfused into HEK293 cells. We propose that the structural features of this AKAP79·PP2B-binding domain may share similarities with other proteins that serve to coordinate PP2B localization and activity.

Original languageEnglish (US)
Pages (from-to)48796-48802
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number50
DOIs
StatePublished - Dec 13 2002

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Calcineurin
Phosphoric Monoester Hydrolases
Protein Kinases
Isoxazoles
Peptides
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
HEK293 Cells
Phosphoprotein Phosphatases
Glutamate Receptors
N-Methylaspartate
Cyclic AMP-Dependent Protein Kinases
Synapses
Protein Kinase C
Signal Transduction
Signal transduction
Proteins
Down-Regulation
Binding Sites
Membranes
Assays

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Mapping the protein phosphatase-2B anchoring site on AKAP79. Binding and inhibition of phosphatase activity are mediated by residues 315-360. / Dell'Acqua, Mark L.; Dodge, Kimberly L.; Tavalin, Steven; Scott, John D.

In: Journal of Biological Chemistry, Vol. 277, No. 50, 13.12.2002, p. 48796-48802.

Research output: Contribution to journalArticle

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AU - Scott, John D.

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