pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.

We report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-...

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Autores principales:	Yunfei Zhang, Qiguang Xie, J Brian Robertson, Carl Hirschie Johnson
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3f36e15c5aa94d69956ca02ceee93b69
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spelling	oai:doaj.org-article:3f36e15c5aa94d69956ca02ceee93b692021-11-18T07:08:49ZpHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.1932-620310.1371/journal.pone.0043072https://doaj.org/article/3f36e15c5aa94d69956ca02ceee93b692012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22905204/?tool=EBIhttps://doaj.org/toc/1932-6203We report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-exhibits pH dependence of its spectral emission in vitro. When expressed in either yeast or mammalian cells, pHlash reports basal pH and cytosolic acidification in vivo. Its spectral ratio response is H(+) specific; neither Ca(++), Mg(++), Na(+), nor K(+) changes the spectral form of its luminescence emission. Moreover, it can be used to image pH in single cells. This is the first BRET-based sensor of H(+) ions, and it should allow the approximation of pH in cytosolic and organellar compartments in applications where current pH probes are inadequate.Yunfei ZhangQiguang XieJ Brian RobertsonCarl Hirschie JohnsonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 8, p e43072 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
description	We report the development of a genetically encodable and ratiometic pH probe named "pHlash" that utilizes Bioluminescence Resonance Energy Transfer (BRET) rather than fluorescence excitation. The pHlash sensor-composed of a donor luciferase that is genetically fused to a Venus fluorophore-exhibits pH dependence of its spectral emission in vitro. When expressed in either yeast or mammalian cells, pHlash reports basal pH and cytosolic acidification in vivo. Its spectral ratio response is H(+) specific; neither Ca(++), Mg(++), Na(+), nor K(+) changes the spectral form of its luminescence emission. Moreover, it can be used to image pH in single cells. This is the first BRET-based sensor of H(+) ions, and it should allow the approximation of pH in cytosolic and organellar compartments in applications where current pH probes are inadequate.
format	article
author	Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson
author_facet	Yunfei Zhang Qiguang Xie J Brian Robertson Carl Hirschie Johnson
author_sort	Yunfei Zhang
title	pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
title_short	pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
title_full	pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
title_fullStr	pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
title_full_unstemmed	pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.
title_sort	phlash: a new genetically encoded and ratiometric luminescence sensor of intracellular ph.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/3f36e15c5aa94d69956ca02ceee93b69
work_keys_str_mv	AT yunfeizhang phlashanewgeneticallyencodedandratiometricluminescencesensorofintracellularph AT qiguangxie phlashanewgeneticallyencodedandratiometricluminescencesensorofintracellularph AT jbrianrobertson phlashanewgeneticallyencodedandratiometricluminescencesensorofintracellularph AT carlhirschiejohnson phlashanewgeneticallyencodedandratiometricluminescencesensorofintracellularph
_version_	1718423848048656384

pHlash: a new genetically encoded and ratiometric luminescence sensor of intracellular pH.

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