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Regulatory phosphorylation of C4 phosphoenolpyruvate carboxylase from Sorghum: An immunological study using specific anti-phosphorylation site-antibodies

Photosynthesis Research volume 43, pages 283–288 (1995)Cite this article

Abstract

A peptide containing the N-terminal phosphorylation site (Ser-8) of Sorghum C4-phospho enolpyruvate carboxylase (PEPC) was synthesized, purified and used to raise an antiserum in rabbits. Affinity-purified IgGs prevented PEPC phosphorylation in a reconstituted in vitro assay and reacted with both the phosphorylated and dephosphorylated forms of either native or denatured PEPC in immunoblotting experiments. Saturation of dephospho-PEPC with these specific IgGs resulted in a marked alteration of its functional and regulatory properties that mimicked phosphorylation of Ser-8. A series of recombinant C4 PEPCs mutated in the N-terminal phosphorylation domain and a C3-like PEPC isozyme from Sorghum behaved similarly to their C4 counterpart with respect to these phosphorylation-site antibodies.

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Abbreviations

PEPC:

phospho enolpyruvate carboxylase

PKA:

catalytic subunit of the cAMP-dependent protein kinase

KLH:

Keyhole Limpet Haemocyanin

IgG:

immunoglobulin G

PEP:

phospho enolpyruvate

SDS-PAGE:

sodium dodecyl sulfate, polyacrylamide gel electrophoresis

MDH:

malate deshydrogenase

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Author information

Affiliations

  1. Institut de Biotechnologie des Plantes, UA CNRS D1128, Université de Paris-Sud, 91405, Orsay Cedex, France

    Valérie Pacquit, Nathalie Giglioli, Claude Crétin, Jean Noel Pierre & Jean Vidal

  2. Laboratorio de Fisiologia Vegetal, Facultad de Biologia, Sevilla, Spain

    Cristina Echevarria

Authors
  1. Valérie Pacquit
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  2. Nathalie Giglioli
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  3. Claude Crétin
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  4. Jean Noel Pierre
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  5. Jean Vidal
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  6. Cristina Echevarria
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Pacquit, V., Giglioli, N., Crétin, C. et al. Regulatory phosphorylation of C4 phosphoenolpyruvate carboxylase from Sorghum: An immunological study using specific anti-phosphorylation site-antibodies. Photosynth Res 43, 283–288 (1995). https://doi.org/10.1007/BF00029941

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  • DOI: https://doi.org/10.1007/BF00029941

Key words

  • antibodies
  • C4 photosynthesis
  • PEPC
  • protein phosphorylation
  • Sorghum