Rice proteomics: a step toward functional analysis of the rice genome.
The technique of proteome analysis with two-dimensional PAGE has the power to monitor global changes that occur in the protein expression of tissues and organisms and/or expression that occurs under stresses. In this study, the catalogues of the rice proteome were constructed, and a functional characterization of some of these proteins was examined. Proteins extracted from tissues of rice and proteins extracted from rice under various kinds of stress were separated by two-dimensional PAGE. An image analyzer was used to reveal a total of 10,589 protein spots on 10 kinds of two-dimensional PAGE gels stained by Coomassie Brilliant Blue. The separated proteins were electroblotted onto a polyvinylidene difluoride membrane, and the N-terminal amino acid sequences of 272 of 905 proteins were determined. The internal amino acid sequences of 633 proteins were determined using a protein sequencer or mass spectrometry after enzyme digestion of the proteins. Finally, a data file of rice proteins that included information on amino acid sequences and sequence homologies was constructed. The major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that turned out to be calreticulin and a gibberellin-binding protein, which is ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice, have functions in the signal transduction pathway. The information thus obtained from the rice proteome will be helpful in predicting the function of the unknown proteins and will aid in their molecular cloning.
Komatsu
Setsuko
S
Department of Molecular Genetics, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan. skomatsu@affrc.go.jp
Konishi
Hirosato
H
Shen
Shihua
S
Yang
Guangxiao
G
D016960:Agrobacterium tumefaciens / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D018745:Genome, Plant; D013058:Mass Spectrometry; D008957:Models, Genetic; D012275:Oryza / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D020543:Proteome
D000596:Amino Acids; D018076:DNA, Complementary; D020543:Proteome