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Phosphorylation of protein Phosphatase-1 isoforms by cdc2-cyclin B in vitro

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Abstract

Protein Phosphatase-1 is phosphorylated in vitro by cdc2-cyclin B (Villa-Moruzzi, FEBS Lett 304: 211-215, 1992). In the present study we show that all the three Phosphatase-1 isoforms, α, γ1, δ, are phosphorylated by cdc2-cyclin B. Phosphorylation is specific for this kinase and involves a C-terminal Thr. This site is most likely Thr 320 in a (shown by others to be phosphorylated also by cdc2-cyclin A). Such Thr is conserved in γ1, δ and in the testis-specific γ2, and is the only Thr that fits the cdc2-consensus sequence in the C-terminal region. Phosphorylation of Phosphatase-1 purified from skeletal muscle, which is a mixture of the a, γ1 and δ isoforms, is up to 0.4 mol/mol and induces 30-35% enzyme inactivation. Following tryptic proteolysis each isoform yields a distinct phosphopeptide map. This is in agreement with the different sequences of the isoforms in the C-terminal regions and may be useful to distinguish the isoforms lts suggest that all the Phosphatase-1 isoforms may be potentially regulated at M-phase.

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  1. Department of Biomedicine, University of Pisa, 56126, Pisa, Italy

    Franca Puntoni & Emma Villa-Moruzzi

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  1. Franca Puntoni
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  2. Emma Villa-Moruzzi
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Puntoni, F., Villa-Moruzzi, E. Phosphorylation of protein Phosphatase-1 isoforms by cdc2-cyclin B in vitro. Mol Cell Biochem 171, 115–120 (1997). https://doi.org/10.1023/A:1006892103306

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