Preparation of Monoclonal Antibodies against NADH : Nitrate Reductase from the Red Alga Porphyra yezoensis : PROTEINS, ENZYMES AND METABOLISM
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概要
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Twenty-two monoclonal antibodies were raised against the native form of nitrate reductase (NR) from Porphyra yezoensis, a marine red alga. All the antibodies were able to bind to NR from P. yezoensis, with resultant inhibition of full (NADH:NR) and/or partial (NADH:FR, NADH:CR, FMNH_2:NR, and MV:NR) enzymatic activity. Fifteen of the antibodies recognized the denatured form of the enzyme. Size-exclusion gel filtration and immunoblotting of the products of the limited proteolysis of NR from P. yezoensis by trypsin or Staphylococcus aureus V8 protease revealed that 2 out of the 1 5 subunit-recognizing antibodies bound to the FAD domain, 6 bound to the heme domain, and 7 bound to the Mo-pterin domain. The products of limited proteolysis of NR from P. yezoensis also revealed that the sites of proteolytic cleavage that encompassed the heme domain were inverted as compared to the analogous sites in NRs of higher plants. Some of the monoclonal antibodies cross-reacted with NRs from plants belonged to different phyla. From three to five of the antibodies bound subunits of NR from multicellular red algae, but failed to bind NRs from unicellular red algae. Three or four of the antibodies cross-reacted with NRs from higher plants, such as tobacco and spinach. One of the antibodies bound NRs from several types of plant, namely, members of Cryptophyta, Chromophyta, and Chlorophyia. All of the monoclonal antibodies that cross-reacted with NRs from plants other than the red algae were specific for the Mo-pterin domain of NR from P. yezoensis.
- 日本植物生理学会の論文
著者
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Saji Hikaru
Regional Environment Division National Institute For Environmental Studies
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KONDO Noriaki
Regional Environment Division, National Institute for Environmental Studies
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Ikawa Tomoyoshi
Institute of Biological Sciences, University of Tsukuba
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Nakamura Yoshiko
Institute of Biological Sciences, University of Tsukuba
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Nakamura Yoshiko
Institute Of Biological Sciences University Of Tsukuba
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Ikawa Tomoyoshi
Institute Of Biological Sciences The University Of Tsukuba
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Kondo Noriaki
Regional And Community Environment Division National Lnstitute For Environmental Studies
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Kondo Noriaki
Regional Environment Division National Institute For Environmental Studies
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