14-3-3 proteins associate with the regulatory phosphorylation site of spinach leaf nitrate reductase in an isoform-specific manner and reduce dephosphorylation of Ser-543 by endogenous protein phosphatases

Markus Bachmann, Joan L. Huber, Gurdeep S. Athwal, Ke Wu, Robert J. Ferl, Steven C. Huber

Research output: Contribution to journalArticle

Abstract

Three lines of evidence indicate that the 14-3-3 proteins that inactivate the phosphorylated form of spinach leaf NADH:nitrate reductase (NR) bind to the enzyme at the regulatory phosphorylation site (Ser-543). First, a phosphorylated synthetic peptide based on the regulatory site can prevent and also reverse the inactivation of phospho-NR caused by 14-3-3 proteins. Second, sequence-specific and phosphorylation-dependent binding of the aforementioned synthetic peptide to the 14-3-3 proteins was demonstrated in vitro. Third, 14-3-3 proteins were required for the ATP-dependent phosphorylation of NR (as assessed by activity measurements) in the presence of NR-kinase and leaf protein phosphatases. Lastly, we demonstrate specificity of recombinant Arabinopsis 14-3-3 isoforms in the interaction with phospho-NR: ω > χ > υ >>> φ, Ψ.

Original languageEnglish (US)
Pages (from-to)26-30
Number of pages5
JournalFEBS Letters
Volume398
Issue number1
DOIs
StatePublished - Nov 25 1996

Fingerprint

14-3-3 Proteins
Nitrate Reductase
Phosphorylation
Spinacia oleracea
Phosphoprotein Phosphatases
Protein Isoforms
Nitrate Reductase (NADH)
Peptides
Phosphotransferases
Adenosine Triphosphate
Enzymes

Keywords

  • 14-3-3 protein
  • Binding site
  • Inhibitor protein
  • Isoform specificity
  • Nitrate reductase
  • Protein phosphatase
  • Regulatory phosphorylation site

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

14-3-3 proteins associate with the regulatory phosphorylation site of spinach leaf nitrate reductase in an isoform-specific manner and reduce dephosphorylation of Ser-543 by endogenous protein phosphatases. / Bachmann, Markus; Huber, Joan L.; Athwal, Gurdeep S.; Wu, Ke; Ferl, Robert J.; Huber, Steven C.

In: FEBS Letters, Vol. 398, No. 1, 25.11.1996, p. 26-30.

Research output: Contribution to journalArticle

Bachmann, Markus ; Huber, Joan L. ; Athwal, Gurdeep S. ; Wu, Ke ; Ferl, Robert J. ; Huber, Steven C. / 14-3-3 proteins associate with the regulatory phosphorylation site of spinach leaf nitrate reductase in an isoform-specific manner and reduce dephosphorylation of Ser-543 by endogenous protein phosphatases. In: FEBS Letters. 1996 ; Vol. 398, No. 1. pp. 26-30.
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abstract = "Three lines of evidence indicate that the 14-3-3 proteins that inactivate the phosphorylated form of spinach leaf NADH:nitrate reductase (NR) bind to the enzyme at the regulatory phosphorylation site (Ser-543). First, a phosphorylated synthetic peptide based on the regulatory site can prevent and also reverse the inactivation of phospho-NR caused by 14-3-3 proteins. Second, sequence-specific and phosphorylation-dependent binding of the aforementioned synthetic peptide to the 14-3-3 proteins was demonstrated in vitro. Third, 14-3-3 proteins were required for the ATP-dependent phosphorylation of NR (as assessed by activity measurements) in the presence of NR-kinase and leaf protein phosphatases. Lastly, we demonstrate specificity of recombinant Arabinopsis 14-3-3 isoforms in the interaction with phospho-NR: ω > χ > υ >>> φ, Ψ.",
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AU - Athwal, Gurdeep S.

AU - Wu, Ke

AU - Ferl, Robert J.

AU - Huber, Steven C.

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