Originally published In Press as doi:10.1074/mcp.M600084-MCP200 on July 6, 2006.
Molecular & Cellular Proteomics 5:2044-2059, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.
Research
Large Scale Identification and Quantitative Profiling of Phosphoproteins Expressed during Seed Filling in Oilseed Rape*,S
Ganesh Kumar Agrawal , ,¶ and
Jay J. Thelen
From the Biochemistry Department, University of Missouri-Columbia, Columbia, Missouri 65211 and Research Laboratory for Agricultural Biotechnology and Biochemistry, P O. Box 8207, Kathmandu, Nepal
Seed filling is a dynamic, temporally regulated phase of seed development that determines the composition of storage reserves in mature seeds. Although the metabolic pathways responsible for storage reserve synthesis such as carbohydrates, oils, and proteins are known, little is known about their regulation. Protein phosphorylation is a ubiquitous form of regulation that influences many aspects of dynamic cellular behavior in plant biology. Here a systematic study has been conducted on five sequential stages (2, 3, 4, 5, and 6 weeks after flowering) of seed development in oilseed rape (Brassica napus L. Reston) to survey the presence and dynamics of phosphoproteins. High resolution two-dimensional gel electrophoresis in combination with a phosphoprotein-specific Pro-Q Diamond phosphoprotein fluorescence stain revealed 300 phosphoprotein spots. Of these, quantitative expression profiles for 234 high quality spots were established, and hierarchical cluster analyses revealed the occurrence of six principal expression trends during seed filling. The identity of 103 spots was determined using LC-MS/MS. The identified spots represented 70 non-redundant phosphoproteins belonging to 10 major functional categories including energy, metabolism, protein destination, and signal transduction. Furthermore phosphorylation within 16 non-redundant phosphoproteins was verified by mapping the phosphorylation sites by LC-MS/MS. Although one of these sites was postulated previously, the remaining sites have not yet been reported in plants. Phosphoprotein data were assembled into a web database. Together this study provides evidence for the presence of a large number of functionally diverse phosphoproteins, including global regulatory factors like 14-3-3 proteins, within developing B. napus seed.
¶ To whom correspondence should be addressed: Biochemistry Dept., University of Missouri-Columbia, 109 Life Sciences Center, Columbia, MO 65211. Tel.: 573-884-5979; Fax: 573-884-9676; E-mail: agrawalg{at}missouri.edu

CiteULike Complore Connotea Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
J. Gao, G. K. Agrawal, J. J. Thelen, and D. Xu
P3DB: a plant protein phosphorylation database
Nucleic Acids Res.,
January 1, 2009;
37(suppl_1):
D960 - D962.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. K. Agrawal, M. Hajduch, K. Graham, and J. J. Thelen
In-Depth Investigation of the Soybean Seed-Filling Proteome and Comparison with a Parallel Study of Rapeseed
Plant Physiology,
September 1, 2008;
148(1):
504 - 518.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. Xu, Y. Xu, and B. Huang
Protein Extraction for Two-Dimensional Gel Electrophoresis of Proteomic Profiling in Turfgrass
Crop Sci.,
July 1, 2008;
48(4):
1608 - 1614.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. M. Lauberth, A. C. Bilyeu, B. A. Firulli, K. L. Kroll, and M. Rauchman
A Phosphomimetic Mutation in the Sall1 Repression Motif Disrupts Recruitment of the Nucleosome Remodeling and Deacetylase Complex and Repression of Gbx2
J. Biol. Chem.,
November 30, 2007;
282(48):
34858 - 34868.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Shin, S. Alvarez, A. Y. Burch, J. M. Jez, and D. P. Schachtman
Phosphoproteomic identification of targets of the Arabidopsis sucrose nonfermenting-like kinase SnRK2.8 reveals a connection to metabolic processes
PNAS,
April 10, 2007;
104(15):
6460 - 6465.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|