A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide

Abstract Genetically encoded temperature indicators (GETIs) allow for real-time measurement of subcellular temperature dynamics in live cells. However, GETIs have suffered from poor temperature sensitivity, which may not be sufficient to resolve small heat production from a biological process. Here,...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Cong Quang Vu, Shun-ichi Fukushima, Tetsuichi Wazawa, Takeharu Nagai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0437663b19484649bddf98f81db09654
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:0437663b19484649bddf98f81db09654
record_format	dspace
spelling	oai:doaj.org-article:0437663b19484649bddf98f81db096542021-12-02T16:28:06ZA highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide10.1038/s41598-021-96049-52045-2322https://doaj.org/article/0437663b19484649bddf98f81db096542021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96049-5https://doaj.org/toc/2045-2322Abstract Genetically encoded temperature indicators (GETIs) allow for real-time measurement of subcellular temperature dynamics in live cells. However, GETIs have suffered from poor temperature sensitivity, which may not be sufficient to resolve small heat production from a biological process. Here, we develop a highly-sensitive GETI, denoted as ELP-TEMP, comprised of a temperature-responsive elastin-like polypeptide (ELP) fused with a cyan fluorescent protein (FP), mTurquoise2 (mT), and a yellow FP, mVenus (mV), as the donor and acceptor, respectively, of Förster resonance energy transfer (FRET). At elevated temperatures, the ELP moiety in ELP-TEMP undergoes a phase transition leading to an increase in the FRET efficiency. In HeLa cells, ELP-TEMP responded to the temperature from 33 to 40 °C with a maximum temperature sensitivity of 45.1 ± 8.1%/°C, which was the highest ever temperature sensitivity among hitherto-developed fluorescent nanothermometers. Although ELP-TEMP showed sensitivity not only to temperature but also to macromolecular crowding and self-concentration, we were able to correct the output of ELP-TEMP to achieve accurate temperature measurements at a subcellular resolution. We successfully applied ELP-TEMP to accurately measure temperature changes in cells induced by a local heat spot, even if the temperature difference was as small as < 1 °C, and to visualize heat production from stimulated Ca2+ influx in live HeLa cells induced by a chemical stimulation. Furthermore, we investigated temperatures in the nucleus and cytoplasm of live HeLa cells and found that their temperatures were almost the same within the temperature resolution of our measurement. Our study would contribute to better understanding of cellular temperature dynamics, and ELP-TEMP would be a useful GETI for the investigation of cell thermobiology.Cong Quang VuShun-ichi FukushimaTetsuichi WazawaTakeharu NagaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cong Quang Vu Shun-ichi Fukushima Tetsuichi Wazawa Takeharu Nagai A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
description	Abstract Genetically encoded temperature indicators (GETIs) allow for real-time measurement of subcellular temperature dynamics in live cells. However, GETIs have suffered from poor temperature sensitivity, which may not be sufficient to resolve small heat production from a biological process. Here, we develop a highly-sensitive GETI, denoted as ELP-TEMP, comprised of a temperature-responsive elastin-like polypeptide (ELP) fused with a cyan fluorescent protein (FP), mTurquoise2 (mT), and a yellow FP, mVenus (mV), as the donor and acceptor, respectively, of Förster resonance energy transfer (FRET). At elevated temperatures, the ELP moiety in ELP-TEMP undergoes a phase transition leading to an increase in the FRET efficiency. In HeLa cells, ELP-TEMP responded to the temperature from 33 to 40 °C with a maximum temperature sensitivity of 45.1 ± 8.1%/°C, which was the highest ever temperature sensitivity among hitherto-developed fluorescent nanothermometers. Although ELP-TEMP showed sensitivity not only to temperature but also to macromolecular crowding and self-concentration, we were able to correct the output of ELP-TEMP to achieve accurate temperature measurements at a subcellular resolution. We successfully applied ELP-TEMP to accurately measure temperature changes in cells induced by a local heat spot, even if the temperature difference was as small as < 1 °C, and to visualize heat production from stimulated Ca2+ influx in live HeLa cells induced by a chemical stimulation. Furthermore, we investigated temperatures in the nucleus and cytoplasm of live HeLa cells and found that their temperatures were almost the same within the temperature resolution of our measurement. Our study would contribute to better understanding of cellular temperature dynamics, and ELP-TEMP would be a useful GETI for the investigation of cell thermobiology.
format	article
author	Cong Quang Vu Shun-ichi Fukushima Tetsuichi Wazawa Takeharu Nagai
author_facet	Cong Quang Vu Shun-ichi Fukushima Tetsuichi Wazawa Takeharu Nagai
author_sort	Cong Quang Vu
title	A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
title_short	A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
title_full	A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
title_fullStr	A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
title_full_unstemmed	A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
title_sort	highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/0437663b19484649bddf98f81db09654
work_keys_str_mv	AT congquangvu ahighlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT shunichifukushima ahighlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT tetsuichiwazawa ahighlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT takeharunagai ahighlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT congquangvu highlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT shunichifukushima highlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT tetsuichiwazawa highlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide AT takeharunagai highlysensitivegeneticallyencodedtemperatureindicatorexploitingatemperatureresponsiveelastinlikepolypeptide
_version_	1718383912648966144

A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide

Ejemplares similares