Molecular recognition moiety and its target biomolecule interact in switching enzyme activity(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
- 論文の詳細を見る
We conjugated a molecular recognition moiety, biotin, with an enzyme site-specifically near to its active site and succeeded in inactivating the enzyme by binding the specific target biomolecule avidin to biotin. Bacterial P450 was used as a model enzyme, which has attracted much attention in several fields. Site-directed mutagenesis was conducted to produce a mutant P450 that could attach biotin site-specifically. The activity of the conjugate decreased markedly to one tenth of that of biotinylated P450 after binding to avidin. Ultraviolet-visible spectroscopy of the carbon monoxide-bound P450, circular dichroism data, and the ratio of the active form to the sum of the active form and the inactive form indicated that this decrease in activity was because of a conformational change in the tertiary structure surrounding the active center after avidin binding, while the secondary structure of P450 remained unchanged.
- 公益社団法人日本生物工学会の論文
- 2013-06-00
著者
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Nagamune Teruyuki
Department Of Bioengineering Center For Nanobio Integration The University Of Tokyo:department Of Ch
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Ohashi Hidenori
Chemical Resources Laboratory, Tokyo Institute of Technology
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Yamaguchi Takeo
Chemical Resources Laboratory, Tokyo Institute of Technology
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TAMAKI Takanori
Chemical Resources Laboratory, Tokyo Institute of Technology
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Yamaguchi Takeo
Chemical Resources Laboratory Tokyo Institute Of Technology
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Ohashi Hidenori
Chemical Resources Laboratory Tokyo Institute Of Technology
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Tamaki Takanori
Chemical Resources Laboratory Tokyo Institute Of Technology
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Hirakawa Hidehiko
Department Of Chemistry And Biotechnology School Of Engineering The University Of Tokyo
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Yamaguchi Satoshi
Department Of Agricultural Chemistry Faculty Of Agriculture Kyushu University
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Nagamune Teruyuki
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo:Department of Bioengineering, School of Engineering, The University of Tokyo
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Oshiba Yuhei
Chemical Resources Laboratory, Tokyo Institute of Technology
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