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Pubmed ID :19376835
Publication Date : //

Large-scale Arabidopsis phosphoproteome profiling reveals novel chloroplast kinase substrates and phosphorylation networks.


We have characterized the phosphoproteome of Arabidopsis (Arabidopsis thaliana) seedlings using high-accuracy mass spectrometry and report the identification of 1,429 phosphoproteins and 3,029 unique phosphopeptides. Among these, 174 proteins were chloroplast phosphoproteins. Motif-X (motif extractor) analysis of the phosphorylation sites in chloroplast proteins identified four significantly enriched kinase motifs, which include casein kinase II (CKII) and proline-directed kinase motifs, as well as two new motifs at the carboxyl terminus of ribosomal proteins. Using the phosphorylation motifs as a footprint for the activity of a specific kinase class, we connected the phosphoproteins with their putative kinases and constructed a chloroplast CKII phosphorylation network. The network topology suggests that CKII is a central regulator of different chloroplast functions. To provide insights into the dynamic regulation of protein phosphorylation, we analyzed the phosphoproteome at the end of day and end of night. The results revealed only minor changes in chloroplast kinase activities and phosphorylation site utilization. A notable exception was ATP synthase beta-subunit, which is found phosphorylated at CKII phosphorylation sites preferentially in the dark. We propose that ATP synthase is regulated in cooperation with 14-3-3 proteins by CKII-mediated phosphorylation of ATP synthase beta-subunit in the dark.

Authors : Reiland Sonja , Messerli Gaëlle , Baerenfaller Katja , Gerrits Bertran , Endler Anne , Grossmann Jonas , Gruissem Wilhelm , Baginsky Sacha ,

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Catalog number Product name Quantity
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AS07 238 Antibody: Toc34 | chloroplast outer envelope membrane translocon complex protein, Immunogen: gel piece with purified GTPase domain of Arabidopsis thaliana that is missing the transmembrane domain with 50
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26-6427-05 3' Phosphorylation 50 nmol scale
26-6427-01 3' Phosphorylation 1 umol scale 1 Each
26-6428-25 Phosphorylation-5' 25 nmol scale
26-6428-05 Phosphorylation-5' 50 nmol scale
26-6659-05 5' phosphorylation DMT ON (CPR-II) 50 nmol scale
26-6428-01 Phosphorylation-5' 1 umol scale 1Each
26-6428-10 Phosphorylation-5' 10 umol scale 1Each
26-6428-02 Phosphorylation-5' 200 nmol scale 1Each
26-6428-03 Phosphorylation-5' 2 umol scale 1Each
26-6428-05 Phosphorylation-5' 50 nmol scale 1Each
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26-6427-10 3' Phosphorylation 10 umol scale 1Each
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26-6428-03 Phosphorylation-5' 2 umol scale
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