The Base Specificities of Tomato Ribonuclease (RNase LE) and Its Asp44 Mutant Enzyme Expressed from Yeast Cells
スポンサーリンク
概要
- 論文の詳細を見る
RNase LE from cultured tomato cells is a member of the RNase T_2 family. It is, however, distinguishable from RNase Rh from Rhizopus niveus, a typical RNase of the RNase T_2 family, by its CD spectrum in the 200-250 nm region. In order to reinvestigate the base specificity of RNase LE and to study the role of Asn44 in RNase LE, which is considered to correspond to the base recognition site Asp51 of RNase Rh, RNase LE, and its Asp mutant at the 44th position were expressed from yeast cells with the same expression system as RNase Rh [K. Ohgi, et al., L Biochem., 109, 776-785 (1991)]. RNase LE with four extra amino acid residues at the 2nd amino acid residue of mature RNase LE and its Asp44 mutant were secreted from yeast cells to give a yield of 10 mg/liter and 0.5 mg/liter culture broth, respectively. The expressed RNase LE (RNase RNAP LE) had the same characteristics as native RNase LE in the CD spectrum and specitic activity. This is the first example of the expression of plant RNase from microbes and in sufficient amount to perform further enzymological research. The base specificity of RNase LE was guanylic acid preferential and that of N44D was changed to a more adenylic acid preference as compared to that of RNase LE. These experiments showed that Asn44 of RNase LE is crucial for base recognition as the case of Asp51 in RNase Rh, and also suggested that the base recognition methanism of RNase LE is very similar to that of RNase Rh.
- 社団法人日本農芸化学会の論文
- 1997-03-23
著者
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Koyama Takashi
Department of Cardiovascular and Respiratory Medicine, Akita University Graduate School of Medicine
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Kobayashi H
National Food Research Institute
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NAKAJIMA ATSUSHI
Department of Gastroenterology, Yokohama City University School of Medicine
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Koyama Takashi
Division Of Life Sciences Graduate School Of Natural Science And Technology Kanazawa University
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Irie Masachika
Department of Microbiology, Hoshi College of Pharmacy
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Ohgi Kazuko
Department of Microbiology, Hoshi College of Pharmacy
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Ohgi K
Department Of Applied Microbiology Faculty Of Pharmaceutical Sciences Hoshi University
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Ohgi Kazuko
Department Of Microbiology Hoshi College Of Pharmacy
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Irie Masachika
Department Of Applied Microbiology Faculty Of Pharmaceutical Sciences Hoshi University
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Irie Masachika
Department Of Microbiology Hoshi College Of Pharmacy
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Kiuchi Hiroshi
Research Development Section Kibun Food Chemifa Co. Ltd.
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KOBAYASHI Hiroko
Department of Internal Medicine II, Fukushima Medical University School of Medicine
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Koyama Takashi
Department Of Microbiology College Of Pharmacy Nihon University
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IWAMA Masanori
Department of Applied Microbiology, Faculty of Pharmaceutical Sciences, Hoshi University
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INOKUCHI Norio
Department of Microbiology, College of Pharmacy, Nihon University
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SHIRATORI Yasuki
Department of Microbiology, College of Pharmacy, Nihon University
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KOCK Margret
Martin-Luther Universitat, Halle-Wittenberg, Biozentrum der Universitat
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LOGGLER Andreas
Martin-Luther Universitat, Halle-Wittenberg, Biozentrum der Universitat
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GLUND Konrad
Martin-Luther Universitat, Halle-Wittenberg, Biozentrum der Universitat(Present address)IFB Halle Gm
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Shiratori Yasuki
Department Of Microbiology College Of Pharmacy Nihon University
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Kock Margret
Martin-luther Universitat Halle-wittenberg Biozentrum Der Universitat
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Loggler Andreas
Martin-luther Universitat Halle-wittenberg Biozentrum Der Universitat:(present Address)ciba-geigy Ag
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Glund Konrad
Martin-luther Universitat Halle-wittenberg Biozentrum Der Universitat(present Address)ifb Halle Gmbh
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Irie M
Hoshi Coll. Pharmacy Tokyo Jpn
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Inokuchi N
Department Of Microbiology College Of Pharmacy Nihon University
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Kobayashi Hiroko
Department Of Gastroenterology And Rheumatology Fukushima Medical University School Of Medicine
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Nakajima Atsushi
Department Of Gastroenterology Yokohama City University Graduate School Of Medicine
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Nakajima Atsushi
Department Of Microbiology College Of Pharmacy Nihon University
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Nakajima Atsushi
Department Of Chemistry Faculty Of Science And Technology Keio University
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Ohgi Kazuko
Department Of Applied Microbiology Faculty Of Pharmaceutical Sciences Hoshi University
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Ohgi K
Department Of Microbiology Hoshi College Of Pharmacy
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Koyama Takashi
Department of Cardiology, Akita University School of Medicine
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Kobayashi Hiroko
Department of Cardiovascular Disease Research, Eisai Tsukuba Research Laboratories
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