Separation and characterization of proteins in rice (Oryza sativa) suspension cultured cells |
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Authors: | Setsuko Komatsu Randeep Rakwal Zhijun Li |
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Affiliation: | (1) Department of Molecular Genetics, National Institute of Agrobiological Resources, 2-1-2, Kannondai, Tsukuba, 305-8602 |
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Abstract: | Proteins extracted from suspension cultured cells of rice were separated by two-dimensional polyacrylamide gel electrophoresis. The separated proteins were electroblotted onto a polyvinylidene difluoride membrane and 103 electroblotted proteins were analyzed. The N-terminal amino-acid sequences of 20 out of 103 proteins were determined in this manner. N-terminal regions of the remaining proteins could not be sequenced and they were inferred to have a blocking group at the N-terminus. Internal amino-acid sequences of 32 proteins were determined by sequence analysis of peptides obtained by Cleveland peptide mapping. The amino-acid sequences determined here were compared with those of known plant and animal proteins. Furthermore, the concanavalin A-peroxidase method was used to determine which of the 103 proteins were glycosylated, and in vitro and in vivo phosphorylation was carried out to identify some of the phosphorylated proteins. Using this experimental approach, we could identify the major proteins involved in growth and development of rice cell suspension cultures and discuss on the physiological function of some of these identified proteins including the calcium binding protein, superoxide dismutase and rice ascorbate peroxidase. This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Graminaceae Oryza sativa protein database rice suspension cultured cells |
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