Publication Information
Niittylä et al., 2007
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Abstract
Mol Cell Proteomics. 2007 Oct;6(10):1711-26. doi: 10.1074/mcp.M700164-MCP200.
Epub 2007 Jun 23.
Temporal analysis of sucrose-induced phosphorylation changes in plasma membrane
proteins of Arabidopsis.
Niittylä T(1), Fuglsang AT, Palmgren MG, Frommer WB, Schulze WX.
Author information:
(1)Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476
Golm, Germany.
Sucrose is the main product of photosynthesis and the most common transport form
of carbon in plants. In addition, sucrose is a compound that serves as a signal
affecting metabolic flux and development. Here we provide first results of
externally induced phosphorylation changes of plasma membrane proteins in
Arabidopsis. In an unbiased approach, seedlings were grown in liquid medium with
sucrose and then depleted of carbon before sucrose was resupplied. Plasma
membranes were purified, and phosphopeptides were enriched and subsequently
analyzed quantitatively by mass spectrometry. In total, 67 phosphopeptides were
identified, most of which were quantified over five time points of sucrose
resupply. Among the identified phosphorylation sites, the well described
phosphorylation site at the C terminus of plasma membrane H(+)-ATPases showed a
relative increase in phosphorylation level in response to sucrose. This
corresponded to a significant increase of proton pumping activity of plasma
membrane vesicles from sucrose-supplied seedlings. A new phosphorylation site
was identified in the plasma membrane H(+)-ATPase AHA1 and/or AHA2. This
phosphorylation site was shown to be crucial for ATPase activity and overrode
regulation via the well known C-terminal phosphorylation site. Novel
phosphorylation sites were identified for both receptor kinases and cytosolic
kinases that showed rapid increases in relative intensities after short times of
sucrose treatment. Seven response classes were identified including
non-responsive, rapid increase (within 3 min), slow increase, and rapid
decrease. Relative quantification of phosphorylation changes by
phosphoproteomics provides a means for identification of fast responses to
external stimuli in plants as a basis for further functional characterization.
DOI: 10.1074/mcp.M700164-MCP200
PMID: 17586839 [Indexed for MEDLINE]
