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j.bmc.2011.06.031.pdf | 1.59 MB | Adobe PDF | 見る/開く |
タイトル: | Construction of dopamine sensors by using fluorescent ribonucleopeptide complexes. |
著者: | Liew, Fong Fong Hasegawa, Tetsuya Fukuda, Masatora Nakata, Eiji https://orcid.org/0000-0001-9564-6805 (unconfirmed) Morii, Takashi https://orcid.org/0000-0003-3663-3267 (unconfirmed) |
著者名の別形: | 森井, 孝 |
キーワード: | In vitro selection Fluorescent sensor Ribonucleopeptide Dopamine Receptor |
発行日: | 1-Aug-2011 |
出版者: | Elsevier Ltd. |
誌名: | Bioorganic & medicinal chemistry |
巻: | 19 |
号: | 15 |
開始ページ: | 4473 |
終了ページ: | 4481 |
抄録: | A facile strategy of stepwise molding of a ribonucleopeptide (RNP) complex affords fluorescent RNP sensors with selective dopamine recognition. In vitro selection of a RNA-derived RNP library, a complex of the Rev peptide and its binding site Rev Responsive Element (RRE) RNA appended with random nucleotides in variable lengths, afforded RNP receptors specific for dopamine. The modular structure of the RNP receptor enables conversion of dopamine-binding RNP receptors to fluorescent dopamine sensors. Application of conditional selection schemes, such as the variation of salt concentrations and application of a counter-selection step by using a competitor ligand norepinephrine resulted in isolation of RNP receptors with defined dopamine-binding characteristics. Increasing the salt condition at the in vitro selection stage afforded RNP receptors with higher dopamine affinity, while addition of norepinephrine in the in vitro selection milieu at the counter-selection step reinforced the selectivity of RNP receptors to dopamine against norepinephrine. Thermodynamic analyses and circular dichroismic studies of the dopamine-RNP complexes suggest that the dopamine-binding RNP with higher selectivity against norepinephrine forms a pre-organized binding pocket and that the dopamine-binding RNP with higher affinity binds dopamine through the induced-fit mechanism. These results indicate that the selection condition controls the ligand-binding mechanism of RNP receptors. |
著作権等: | © 2011 Elsevier Ltd. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 This is not the published version. Please cite only the published version. |
URI: | http://hdl.handle.net/2433/147155 |
DOI(出版社版): | 10.1016/j.bmc.2011.06.031 |
PubMed ID: | 21742507 |
出現コレクション: | 学術雑誌掲載論文等 |
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