Protein phosphatase 1 catalytic subunit isoforms from alfalfa: Biochemical characterization and cDNA cloning

Emese Vissi, Éva Csordás Tóth, Izabella Kovács, Zoltán Magyar, Gábor V. Horváth, Péter Bagossi, Pál Gergely, Dénes Dudits, Viktor Dombrádi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The catalytic subunit of protein phosphatase 1 (PP1c) was purified from an alfalfa (Medicago sativa) microcallus cell culture. The preparation was inhibited by rabbit muscle inhibitor-2 and okadaic acid and had a molecular mass of 35 kDa. Five distinct cDNAs termed MsPP1α, -β, -γ, -δ, and -ε were cloned from a M. sativa somatic embryo library. MsPP1α was identical to a cDNA reported earlier [A. Pay, M. Pirck, L. Bogre, H. Hirt, and E. Heberle- Bors Mol. Gen. Genet. 244, 176-182, 1994], while the others represented novel isoforms encoded by separate genes. The predicted amino acid sequences of MsPP1α, -β, -γ, -δ, and -ε were highly similar to each other and to other known PP1c sequences. The GST-MsPP1β fusion protein expressed in Escherichia coli was catalytically active and was inhibited by inhibitor-2 and okadaic acid. Affinity-purified polyclonal MsPP1 antipeptide antibody detected a protein of 36 kDa in crude cell extracts. These results proved that the cDNA clone encoded an active PP1c which was very similar to the purified enzyme. The mRNA and protein concentrations of PP1c as well as the specific activity of protein phosphatase 1 did not change during the cell cycle in a synchronized alfalfa cell culture. On the other hand, the isoforms exhibited different steady-state mRNA levels in different plant organs suggesting tissue-specific functions.

Original languageEnglish
Pages (from-to)206-214
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume360
Issue number2
DOIs
Publication statusPublished - Dec 15 1998

Keywords

  • Cell cycle
  • Gene expression
  • Medicago sativa
  • Protein phosphatase 1
  • Protein purification
  • cDNA sequencing

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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