, Volume 233, Issue 2, pp 383–392 | Cite as

Large-scale analysis of phosphorylated proteins in maize leaf

  • Ying-Dong Bi
  • Hong-Xia Wang
  • Tian-Cong Lu
  • Xiao-hui Li
  • Zhuo Shen
  • Yi-Bo Chen
  • Bai-Chen WangEmail author
Original Article


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.


Maize NanoLC–MS/MS Phosphorylation 



Titanium dioxide


Nano liquid chromatography-tandem mass spectrometry


Phosphoenolpyruvate carboxylase


Nitrate reductase


Histone deacetylases



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).

Supplementary material

425_2010_1291_MOESM1_ESM.doc (398 kb)
Supplementary material 1 (DOC 397 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Ying-Dong Bi
    • 1
    • 2
  • Hong-Xia Wang
    • 3
  • Tian-Cong Lu
    • 1
  • Xiao-hui Li
    • 4
  • Zhuo Shen
    • 1
  • Yi-Bo Chen
    • 1
  • Bai-Chen Wang
    • 1
    Email author
  1. 1.Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education and School of ForestryNortheast Forestry UniversityHarbinPeople’s Republic of China
  2. 2.Research Institute of Crop CultivationHeilongjiang Academy of Agricultural SciencesHarbinChina
  3. 3.National Center of Biomedical AnalysisBeijingChina
  4. 4.Center of Agri-BiotechnologyJilin Academy of Agricultural SciencesChangchunChina

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