Abstract
Proteomic analyses of the β subunit of the plastid ATP synthase of barley (Hordeum vulgare L.) revealed that mature protein was not carboxy terminus processed and suggested the correction of the 274 codon (GAT to AAT) in the data bank that was confirmed by DNA sequencing. Six isoforms of the ATP synthase β subunit with pI ranging from 4.95 to 5.14 were resolved by two-dimensional electrophoresis (2-DE). Mass spectrometry analyses indicated that the six isoforms differ in their phosphorylation degree, which was confirmed by the disappearance of more acidic forms after incubation with the protein phosphatase calcineurin. Six Ser and/or Thr were detected as phosphorylated, among them the conserved Thr-179 that is also phosphorylated in the β subunit of human mitochondria. The results are discussed in relation with the proposed regulation of the ATP synthase by phosphorylation and 14-3-3 proteins.
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Abbreviations
- 2-DE:
-
two-dimensional electrophoresis
- ESI-MS/MS:
-
electro spray ionisation tandem mass spectrometry
- IEF:
-
isoelectric focusing
- MALDI-TOF:
-
matrix-assisted laser desorption/ionisation time-of-flight
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Acknowledgements
This work was supported by a Grant from the Spanish DGICYT (BMC2003-01261). G. del Riego is recipient of a fellowship of the Spanish MEC. We thank P.H. Serrot for his help in performing DNA sequencing and for English corrections.
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del Riego, G., Casano, L.M., Martín, M. et al. Multiple phosphorylation sites in the β subunit of thylakoid ATP synthase. Photosynth Res 89, 11–18 (2006). https://doi.org/10.1007/s11120-006-9078-4
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DOI: https://doi.org/10.1007/s11120-006-9078-4