Regulatory phosphorylation of C4 phosphoenolpyruvate carboxylase from Sorghum: An immunological study using specific anti-phosphorylation site-antibodies |
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Authors: | Valérie Pacquit Nathalie Giglioli Claude Crétin Jean Noel Pierre Jean Vidal Cristina Echevarria |
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Institution: | (1) Institut de Biotechnologie des Plantes, UA CNRS D1128, Université de Paris-Sud, 91405 Orsay Cedex, France;(2) Laboratorio de Fisiologia Vegetal, Facultad de Biologia, Sevilla, Spain |
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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.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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Keywords: | antibodies C4 photosynthesis PEPC protein phosphorylation Sorghum |
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