Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes

Summary: The mitochondrial calcium uniporter, which mediates mitochondrial Ca2+ uptake, regulates key cellular functions, including intracellular Ca2+ signaling, cell-fate determination, and mitochondrial bioenergetics. Here, we describe two complementary strategies to quantify the uniporter’s trans...

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Autores principales:	Madison X. Rodriguez, Anna M. Van Keuren, Ming-Feng Tsai
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Biophysics Cell Biology Cell-based Assays Molecular Biology Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/ce787d4030db46d59014f17d1360c5dc
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spelling	oai:doaj.org-article:ce787d4030db46d59014f17d1360c5dc2021-11-26T04:40:42ZQuantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes2666-166710.1016/j.xpro.2021.100979https://doaj.org/article/ce787d4030db46d59014f17d1360c5dc2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006857https://doaj.org/toc/2666-1667Summary: The mitochondrial calcium uniporter, which mediates mitochondrial Ca2+ uptake, regulates key cellular functions, including intracellular Ca2+ signaling, cell-fate determination, and mitochondrial bioenergetics. Here, we describe two complementary strategies to quantify the uniporter’s transport activity. First, we detail a mitochondrial Ca2+ radionuclide uptake assay in cultured cell lines. Second, we describe electrophysiological recordings of the uniporter expressed in Xenopus oocytes. These approaches enable a detailed kinetic analysis of the uniporter to link its molecular properties to physiological functions.For complete details on the use and execution of this protocol, please refer to Tsai and Tsai (2018) and Phillips et al. (2019).Madison X. RodriguezAnna M. Van KeurenMing-Feng TsaiElsevierarticleBiophysicsCell BiologyCell-based AssaysMolecular BiologyScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100979- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biophysics Cell Biology Cell-based Assays Molecular Biology Science (General) Q1-390
spellingShingle	Biophysics Cell Biology Cell-based Assays Molecular Biology Science (General) Q1-390 Madison X. Rodriguez Anna M. Van Keuren Ming-Feng Tsai Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
description	Summary: The mitochondrial calcium uniporter, which mediates mitochondrial Ca2+ uptake, regulates key cellular functions, including intracellular Ca2+ signaling, cell-fate determination, and mitochondrial bioenergetics. Here, we describe two complementary strategies to quantify the uniporter’s transport activity. First, we detail a mitochondrial Ca2+ radionuclide uptake assay in cultured cell lines. Second, we describe electrophysiological recordings of the uniporter expressed in Xenopus oocytes. These approaches enable a detailed kinetic analysis of the uniporter to link its molecular properties to physiological functions.For complete details on the use and execution of this protocol, please refer to Tsai and Tsai (2018) and Phillips et al. (2019).
format	article
author	Madison X. Rodriguez Anna M. Van Keuren Ming-Feng Tsai
author_facet	Madison X. Rodriguez Anna M. Van Keuren Ming-Feng Tsai
author_sort	Madison X. Rodriguez
title	Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
title_short	Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
title_full	Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
title_fullStr	Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
title_full_unstemmed	Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes
title_sort	quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or xenopus oocytes
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/ce787d4030db46d59014f17d1360c5dc
work_keys_str_mv	AT madisonxrodriguez quantitativeassaystomeasurethetransportactivityofthemitochondrialcalciumuniporterincelllinesorxenopusoocytes AT annamvankeuren quantitativeassaystomeasurethetransportactivityofthemitochondrialcalciumuniporterincelllinesorxenopusoocytes AT mingfengtsai quantitativeassaystomeasurethetransportactivityofthemitochondrialcalciumuniporterincelllinesorxenopusoocytes
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Quantitative assays to measure the transport activity of the mitochondrial calcium uniporter in cell lines or Xenopus oocytes

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