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Large-scale analysis of phosphorylated proteins in maize leaf

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Abstract

Phosphorylation is an ubiquitous regulatory mechanism governing the activity, subcellular localization, and intermolecular interactions of proteins. To identify a broad range of phosphoproteins from Zea mays, we enriched phosphopeptides from Zea mays leaves using titanium dioxide microcolumns and then extensively fractionated and identified the phosphopeptides by mass spectrometry. A total of 165 unique phosphorylation sites with a putative role in biological processes were identified in 125 phosphoproteins. Most of these proteins are involved in metabolism, including carbohydrate and protein metabolism. We identified novel phosphorylation sites on translation initiation factors, splicing factors, nucleolar RNA helicases, and chromatin-remodeling proteins such as histone deacetylases. Intriguingly, we also identified phosphorylation sites on several proteins associated with photosynthesis, and we speculate that these sites may be involved in carbohydrate metabolism or electron transport. Among these phosphoproteins, phosphoenolpyruvate carboxylase and NADH: nitrate reductase (NR) which catalyzes the rate-limiting and regulated step in the pathway of inorganic nitrogen assimilation were identified. A conserved phosphorylation site was found in the cytochrome b5 heme-binding domain of NADH: nitrate reductase, suggesting that NADH: nitrate reductase is phosphorylated by the same protein kinase or highly related kinases. These data demonstrate that the pathways that regulate diverse processes in plants are major targets of phosphorylation.

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Abbreviations

TiO2 :

Titanium dioxide

NanoLC–MS/MS:

Nano liquid chromatography-tandem mass spectrometry

PEPC:

Phosphoenolpyruvate carboxylase

NADH:

Nitrate reductase

HDACs:

Histone deacetylases

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Acknowledgments

We are grateful for financial support from the Fundamental Research Funds for the Central Universities (DL09EA01), Key Project of Science and Technology of the Ministry of Education of the People’s Republic of China (Grant 108048), and the Natural Sciences Foundation of Heilongjiang Province (Grant C200930).

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Authors and Affiliations

  1. Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education and School of Forestry, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, People’s Republic of China

    Ying-Dong Bi, Tian-Cong Lu, Zhuo Shen, Yi-Bo Chen & Bai-Chen Wang

  2. Research Institute of Crop Cultivation, Heilongjiang Academy of Agricultural Sciences, 368 Xuefu Road, Harbin, 150086, China

    Ying-Dong Bi

  3. National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, China

    Hong-Xia Wang

  4. Center of Agri-Biotechnology, Jilin Academy of Agricultural Sciences, 1363 Caiyu Road, Changchun, 130033, China

    Xiao-hui Li

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  1. Ying-Dong Bi
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  2. Hong-Xia Wang
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  3. Tian-Cong Lu
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  4. Xiao-hui Li
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  5. Zhuo Shen
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Correspondence to Bai-Chen Wang.

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Y.-D. Bi, H.-X. Wang, and T.-C. Lu have contributed equally to the paper.

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Bi, YD., Wang, HX., Lu, TC. et al. Large-scale analysis of phosphorylated proteins in maize leaf. Planta 233, 383–392 (2011). https://doi.org/10.1007/s00425-010-1291-x

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