Regulated interaction of protein phosphatase 1 and protein phosphatase 2A with phospholipase C-related but catalytically inactive protein

Goro Sugiyama, Hiroshi Takeuchi, Koki Nagano, Jing Gao, Yukiko Ohyama, Yoshihide Mori, Masato Hirata

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Protein phosphatase 1 (PP1) and protein phosphatase 2A (PP2A) are major members of the protein serine/threonine phosphatase families. We have identified PP1 and PP2A as interacting partners of PRIP (phospholipase C-related but catalytically inactive protein), a protein isolated in our laboratory. We first investigated the interaction of PRIP with two phosphatases, using purified recombinant proteins. PRIP immobilized on beads pulled down the catalytic subunits of both PP1 and PP2A, indicating that the interactions were in a direct manner, and the binding of PP1 and the binding of PP2A to PRIP were mutually exclusive. Site-directed mutagenesis experiments revealed that the binding sites for PP1 and PP2A on PRIP were not identical, but similar. Phosphorylation of PRIP by protein kinase A (PKA) resulted in the weakened binding of PP1, but not PP2A. Rather, the dissociation of PP1 from PRIP by phosphorylation accompanied the strengthened binding of PP2A in in vitro experiments. This regulation of binding of PP1 and PP2A to PRIP by PKA-dependent phosphorylation was also observed in living cells treated with forskolin or isoproterenol. These results suggested that PRIP directly interacts with the catalytic subunits of two distinct phosphatases in a mutually exclusive manner and the interactions are regulated by phosphorylation, thus functioning as a scaffold to regulate the activities and subcellular localizations of both PP1 and PP2A in phospho-dependent cellular signaling.

Original languageEnglish
Pages (from-to)3394-3403
Number of pages10
JournalBiochemistry
Volume51
Issue number16
DOIs
Publication statusPublished - Apr 24 2012

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Protein Phosphatase 1
Protein Phosphatase 2
Type C Phospholipases
Carrier Proteins
Phosphorylation
Proteins
Cyclic AMP-Dependent Protein Kinases
Phosphoric Monoester Hydrolases
Cell signaling
Mutagenesis
Phosphoprotein Phosphatases
Colforsin
Site-Directed Mutagenesis
Isoproterenol
Recombinant Proteins
Scaffolds
Catalytic Domain
Experiments
Binding Sites
Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Regulated interaction of protein phosphatase 1 and protein phosphatase 2A with phospholipase C-related but catalytically inactive protein. / Sugiyama, Goro; Takeuchi, Hiroshi; Nagano, Koki; Gao, Jing; Ohyama, Yukiko; Mori, Yoshihide; Hirata, Masato.

In: Biochemistry, Vol. 51, No. 16, 24.04.2012, p. 3394-3403.

Research output: Contribution to journalArticle

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AU - Hirata, Masato

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