Proteomics of the rice cell: systematic identification of the protein populations in subcellular compartments

被引:89
作者
Tanaka, N
Fujita, M
Handa, H
Murayama, S
Uemura, M
Kawamura, Y
Mitsui, T
Mikami, S
Tozawa, Y
Yoshinaga, T
Komatsu, S [1 ]
机构
[1] Natl Inst Agroenvironm Sci, Dept Mol Genet, Tsukuba, Ibaraki 3058602, Japan
[2] Natl Agr Res Ctr Hokkaido Reg, Sapporo, Hokkaido 0628555, Japan
[3] Iwate Univ, Fac Agr, Cryobiosyst Res Ctr, Morioka, Iwate 0208550, Japan
[4] Niigata Univ, Grad Sch Sci & Technol, Niigata 9502181, Japan
[5] Mitsubishi Kagaku Inst Life Sci, Translat Res Dept, Yokohama, Kanagawa 2278502, Japan
关键词
proteome; Oryza saliva; two-dimensional electrophoresis; mass spectrometry;
D O I
10.1007/s00438-004-1002-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Despite recent progress in sequencing the complete genome of rice (Oryza saliva), the proteome of this species remains poorly understood. To extend our knowledge of the rice proteome, the subcellular compartments, which include plasma membranes (PM), vacuolar membranes (VM), Golgi membranes (GM), mitochondria (MT), and chloroplasts (CP), were purified from rice seedlings and cultured suspension cells. The proteins of each of these compartments were then systematically analyzed using two-dimensional (2D) electrophoresis, mass spectrometry, and Edman sequencing, followed by database searching. In all, 58 of the 464 spots detected by 2D electrophoresis in PM, 43 of the 141 spots in VM, 46 of the 361 spots in GM, 146 in the 672 spots in MT, and 89 of the 252 spots in CP could be identified by this procedure. The characterized proteins were found to be involved in various processes, such as respiration and the citric acid cycle in MT; photosynthesis and ATP synthesis in CP; and antiftingal defense and signal systems in the membranes. Edman degradation revealed that 60-98% of N-terminal sequences were blocked, and the ratios of blocked to unblocked proteins in the proteomes of the various subcellular compartments differed. The data on the proteomes of subcellular compartments in rice will be valuable for resolving questions in functional genomics as well as for genome-wide exploration of plant function.
引用
收藏
页码:566 / 576
页数:11
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