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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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