Immunochemical Analysis of Multiple Subunit Polypeptides of Glutamine Synthetase in Methionine Sulfoximine-Sensitive and Tolerant Tobacco Cell Cultures
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概要
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A methionine sulfoximine (MSX) tolerant cell line of tobacco (Nicotiana tabacum L. cv. Xanthi) cells was selected by culturing the wild-type cells in suspension media in the presence of MSX at a step-wise increase in its concentration (0.3 to 5 μM). Fifty per cent inhibition of growth occur-red at 0.18 μM MSX for the wild-type cells whereas 4.65 μM was required for the tolerant cells. The tolerant cells possessed about 1.5-fold increase in glutamine synthetase (GO) activity. Kinetic experiments showed that an inhibitor constant for MSX was identical between GO isolated from these two cell types. Subunit polypeptides of GS in both cell types were analyzed with an immunoblotting method by using polyclonal antibody raised against a chloroplastic GS in spinach. A single polypeptide (41 kDa) was recognized by the antibody in wild-type cells, whereas two predominant polypeptides of 41 and 40 kDa were seen in the MSX tolerant cells. When the GS subunit polypeptides in the wild-type cells were examined with two-dimensional gel electrophoresis, two major and four minor polypeptides associating distinct charge at 41 kDa were detected. The extract from the MSX-tolerant cells had the same set of polypeptides at 41 kDa and in addition two major and some minor spots at 40 kDa. These results indicate that 1) tobacco GS is consisted of heterogeneous subunit polypeptides in surface charge and 2) MSX causes formation of additional multiple 40 kDa polypeptides which may be related to the tolerant nature of the selected cell line.
著者
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Yamaya Tomoyuki
Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University
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Yamaya T
Tohoku Univ. Sendai Jpn
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Yamaya T
Institute For Agricultural And Biological Sciences. Okayama University
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Yamaya Tomoyuki
Laboratory Of Plant Cell Engineering Department Of Agricultural Chemistry Faculty Of Agriculture Toh
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Yamaya Tomoyuki
Laboratory Of Plant Cell Biochemistry Department Of Applied Plant Science Division Of Life Science G
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Ojima Kunihiko
Laboratory of Plant Nutrition and Laboratory of Cell Biochemistry, Department of Applied Biological
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KAMACHI Kazunari
Laboratory of Plant Nutrition, Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku
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Kamachi Kazunari
Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University
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Kamachi K
Department Of Bacterial Pathogenesis And Infection Control National Institute Of Infectious Diseases
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Yoneyama T
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni
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Ojima K
Laboratory Of Plant Nutrition Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku Uni
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Ojima Kunihiko
Laboratory Of Plant Nutrition And Laboratory Of Cell Biochemistry Department Of Applied Biological C
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Ishida Hirohiko
Laboratory of Plant Nutrition, Department of Agricultural Chemistry, Faculty of Agriculture,Tohoku U
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Ishida Hirohiko
Laboratory Of Plant Nutrition Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku Uni
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