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タイトル: | Glycine insertion makes yellow fluorescent protein sensitive to hydrostatic pressure. |
著者: | Watanabe, Tomonobu M Imada, Katsumi Yoshizawa, Keiko Nishiyama, Masayoshi Kato, Chiaki Abe, Fumiyoshi Morikawa, Takamitsu J Kinoshita, Miki Fujita, Hideaki Yanagida, Toshio |
著者名の別形: | 西山, 雅祥 |
発行日: | 27-Aug-2013 |
出版者: | Public Library of Science |
誌名: | PloS one |
巻: | 8 |
号: | 8 |
論文番号: | e73212 |
抄録: | Fluorescent protein-based indicators for intracellular environment conditions such as pH and ion concentrations are commonly used to study the status and dynamics of living cells. Despite being an important factor in many biological processes, the development of an indicator for the physicochemical state of water, such as pressure, viscosity and temperature, however, has been neglected. We here found a novel mutation that dramatically enhances the pressure dependency of the yellow fluorescent protein (YFP) by inserting several glycines into it. The crystal structure of the mutant showed that the tyrosine near the chromophore flipped toward the outside of the β-can structure, resulting in the entry of a few water molecules near the chromophore. In response to changes in hydrostatic pressure, a spectrum shift and an intensity change of the fluorescence were observed. By measuring the fluorescence of the YFP mutant, we succeeded in measuring the intracellular pressure change in living cell. This study shows a new strategy of design to engineer fluorescent protein indicators to sense hydrostatic pressure. |
著作権等: | © 2013 Watanabe et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
URI: | http://hdl.handle.net/2433/178907 |
DOI(出版社版): | 10.1371/journal.pone.0073212 |
PubMed ID: | 24014139 |
出現コレクション: | 学術雑誌掲載論文等 |
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