Preparation of Neocarzinostatin Apoprotein Mutants and the Randomized Library on the Chromophore-Binding Cavity(Miscellaneous)
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概要
- 論文の詳細を見る
W39F, F52Y, S98G, S98A, and S98C mutants of the neocarzinostatin apoprotein (apo-NCS) were newly prepared and investigated their physicochemical properties. The circular dichroism (CD) spectra of F78W, F52Y, S98A, S98G, S98C were superimposable with that of wild type 1R49 protein although the minor spectral change seemed to be in the ellipticity of W39F. The results suggest that position 52, 78, and 98 involving natural chromophore binding do not play a major role in the inducing overall structural changes of the protein. Conversely, the position 39 would be affected slightly. Ethidium bromide (EtdBr) binding to mutants was also evaluated by the monitoring of total fluorescence intensity and fluorescence polarization (FP). The observed dissociation constant in the FP study was 4.4μM for wild type, 2.2μM for S98A, 1.3μM for S98G, 9.7μM for S98C, respectively. When S98G and F52Y, the calculated maximum change of the total fluorescence intensity was increased, suggesting that the EtdBr binding to S98G or F52Y were slightly improved compared with the wild type. Then, a total of 14 amino acids randomly substituted phage displayed library of apo-NCS was successfully prepared, because substitution of the amino acid structured the chromophore-binding cavity were not change the overall structural features. The phages which bound glycyrrhetic acid conjugated bovine serum albumin were enriched from this library using phage display technique as the pilot experiments. Although more precision investigation still needs, it should be possible to select variants that have new functions not found in nature.
- 公益社団法人日本薬学会の論文
- 2006-05-01
著者
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MIZUGAKI Michinao
Department of Clinical Pharmaceutics, Tohoku Pharmaceutical University
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HISHINUMA TAKANORI
Division of Pharmacotherapy, Graduate School of Pharmaceutical Sciences, Tohoku University
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Goto J
Tohoku Univ. Sendai Jpn
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Goto J
The Ion Engineering Research Institute Corporation
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Yamaguchi Hiroaki
Division Of Prosthetics Department Of Oral And Maxillofacial Rehabilitation Kanagawa Dental College
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Goto J
Division Of Clinical Pharmacy Graduate School Of Pharmaceutical Sciences Tohoku University
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GOTO Junichi
Division of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Tohoku University
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TOMIOKA Yoshihisa
Division of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Tohoku University
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KISARA Shigeki
Division of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Tohoku University
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YOSHIZAWA Saki
Division of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Tohoku University
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OZAWA Mika
Laboratory of Pharmacotherapeutics, Department of Pharmaceutical Sciences, Josai International Unive
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SUZUKI Naoto
Laboratory of Pharmacotherapeutics, Department of Pharmaceutical Sciences, Josai International Unive
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Mizugaki Michinao
Department Of Clinical Pharmaceutics Tohoku Pharmaceutical University
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Mizugaki Michinao
Dept. Pharmaceutical Sciences Akita University
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Hayashi T
Department Of Pharmacy Fujita Health University Hospital
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Suzuki N
Laboratory Of Pharmacotherapeutics Department Of Pharmaceutical Sciences Josai International Univers
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Nozaki Seishiro
Department Of Pharmaceutical Sciences Tohoku University Hospital And Division Of Clinical Pharmacy G
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Nozaki Seishiro
Department Of Pharmacy Ohta Nishinouchi Hospital
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Ozawa Mika
Laboratory Of Pharmacotherapeutics Department Of Pharmaceutical Sciences Josai International Univers
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Tomioka Yoshihisa
Division Of Clinical Pharmacy Graduate School Of Pharmaceutical Sciences Tohoku University:laborator
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Hishinuma Takanori
Division Of Pharmacotherapy Graduate School Of Pharmaceutical Sciences Tohoku University
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Yoshizawa Saki
Division Of Clinical Pharmacy Graduate School Of Pharmaceutical Sciences Tohoku University
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Kisara Shigeki
Division Of Clinical Pharmacy Graduate School Of Pharmaceutical Sciences Tohoku University
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Mizugaki M
Department Of Clinical Pharmaceutics Tohoku Pharmaceutical University
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Mizugaki M
Department Of Pharmaceutical Sciences Tohoku University Hospital
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Hishinuma T
Division Of Clinical Pharmacy Graduate School Of Pharmaceutical Sciences Tohoku University
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Goto Junichi
Department of Pharmaceutical Sciences, Tohoku University Hospital
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Yamaguchi Hiroaki
Department of Pharmaceutical Sciences, Tohoku University Hospital
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