Enzymatic Properties of Serine 93 Mutants of RNase Rh from Rhizopus niveus. : A Trial to Alter the Base Preference of RNase Rh(Biochemistry)
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概要
- 論文の詳細を見る
In order to investigate the effects of mutation of Ser93, a component of base recognition site (B2 site) of a base non-specific RNase from Rhizopus niveus, we prepared 10 mutant enzymes at this position, S93A, S93V, S93F, S93T, S93G, S93D, S93N, S93E, S93Q and S93R, and their enzymatic activities towards RNA and 16 dinucleoside phosphates were measured. Enzymatic activities of the mutant enzymes towards RNA were between 3.5-75% of the native enzyme. From the rates of hydrolysis of 16 dinucleoside phosphates by the mutant enzymes, we estimated the base preference of B1 and B2 base recognition sites. The results indicated that mutation of Ser93 to Phe, Thr, Glu. Gln and Arg caused the B2 site of the enzymes to more cytosine base preference and Asp and Asn substitution caused more uracil base preference. The results suggested that we are able to construct an enzyme that preferentially cleaves internucleotidic linkage at the 5'-side of cytidine or uridine. The results seem able to convert a base non-specific RNase to a base specific one.
- 公益社団法人日本薬学会の論文
- 2005-10-01
著者
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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 M
Department Of Applied Microbiology Faculty Of Pharmaceutical Sciences Hoshi University
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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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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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SANDA Akihiro
Department of Public Health, Azabu University
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Sanda A
Department Of Hygienic Technology College Of Environmental Health Azabu University
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Irie M
Hoshi Coll. Pharmacy Tokyo Jpn
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Iwama M
Department Of Microbiology Faculty Of Pharmaceutical Sciences Ohu-university
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Inokuchi N
Department Of Microbiology College Of Pharmacy Nihon 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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Sanda Akihiro
Department of Hygienic Technology, College of Environmental Health, Azabu University
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