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Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure
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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3754940/ https://www.ncbi.nlm.nih.gov/pubmed/24014139 http://dx.doi.org/10.1371/journal.pone.0073212 |
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author | Watanabe, Tomonobu M. Imada, Katsumi Yoshizawa, Keiko Nishiyama, Masayoshi Kato, Chiaki Abe, Fumiyoshi Morikawa, Takamitsu J. Kinoshita, Miki Fujita, Hideaki Yanagida, Toshio |
author_facet | Watanabe, Tomonobu M. Imada, Katsumi Yoshizawa, Keiko Nishiyama, Masayoshi Kato, Chiaki Abe, Fumiyoshi Morikawa, Takamitsu J. Kinoshita, Miki Fujita, Hideaki Yanagida, Toshio |
author_sort | Watanabe, Tomonobu M. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3754940 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37549402013-09-06 Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure Watanabe, Tomonobu M. Imada, Katsumi Yoshizawa, Keiko Nishiyama, Masayoshi Kato, Chiaki Abe, Fumiyoshi Morikawa, Takamitsu J. Kinoshita, Miki Fujita, Hideaki Yanagida, Toshio PLoS One Research Article 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. Public Library of Science 2013-08-27 /pmc/articles/PMC3754940/ /pubmed/24014139 http://dx.doi.org/10.1371/journal.pone.0073212 Text en © 2013 Watanabe et al http://creativecommons.org/licenses/by/4.0/ 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 properly credited. |
spellingShingle | Research Article Watanabe, Tomonobu M. Imada, Katsumi Yoshizawa, Keiko Nishiyama, Masayoshi Kato, Chiaki Abe, Fumiyoshi Morikawa, Takamitsu J. Kinoshita, Miki Fujita, Hideaki Yanagida, Toshio Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title | Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title_full | Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title_fullStr | Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title_full_unstemmed | Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title_short | Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure |
title_sort | glycine insertion makes yellow fluorescent protein sensitive to hydrostatic pressure |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3754940/ https://www.ncbi.nlm.nih.gov/pubmed/24014139 http://dx.doi.org/10.1371/journal.pone.0073212 |
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