Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells

Summary: Endocytic internalization of extracellular proteins plays roles in signaling, nutrient uptake, immunity, and extracellular protein quality control. However, there are few protocols for analyzing the lysosomal degradation of extracellular protein. Here, we purified secreted proteins fused wi...

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Autores principales:	Ayaka Tomihari, Momoka Chiba, Akira Matsuura, Eisuke Itakura
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Cell Biology Flow Cytometry/Mass Cytometry Cell-based Assays Molecular/Chemical Probes Protein expression and purification Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/73c2926b026747bab84edd04ce386c97
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spelling	oai:doaj.org-article:73c2926b026747bab84edd04ce386c972021-11-26T04:40:40ZProtocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells2666-166710.1016/j.xpro.2021.100975https://doaj.org/article/73c2926b026747bab84edd04ce386c972021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S266616672100681Xhttps://doaj.org/toc/2666-1667Summary: Endocytic internalization of extracellular proteins plays roles in signaling, nutrient uptake, immunity, and extracellular protein quality control. However, there are few protocols for analyzing the lysosomal degradation of extracellular protein. Here, we purified secreted proteins fused with pH-sensitive GFP and acid- and protease-resistant RFP from mammalian cells and describe an internalization assay for mammalian cells. This protocol enables quantification of cellular uptake and lysosomal degradation of protein-of-interest (POI) via cell biological and biochemical analyses.For full details on the use and execution of this protocol, please refer to Itakura et al. (2020).Ayaka TomihariMomoka ChibaAkira MatsuuraEisuke ItakuraElsevierarticleCell BiologyFlow Cytometry/Mass CytometryCell-based AssaysMolecular/Chemical ProbesProtein expression and purificationScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100975- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cell Biology Flow Cytometry/Mass Cytometry Cell-based Assays Molecular/Chemical Probes Protein expression and purification Science (General) Q1-390
spellingShingle	Cell Biology Flow Cytometry/Mass Cytometry Cell-based Assays Molecular/Chemical Probes Protein expression and purification Science (General) Q1-390 Ayaka Tomihari Momoka Chiba Akira Matsuura Eisuke Itakura Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
description	Summary: Endocytic internalization of extracellular proteins plays roles in signaling, nutrient uptake, immunity, and extracellular protein quality control. However, there are few protocols for analyzing the lysosomal degradation of extracellular protein. Here, we purified secreted proteins fused with pH-sensitive GFP and acid- and protease-resistant RFP from mammalian cells and describe an internalization assay for mammalian cells. This protocol enables quantification of cellular uptake and lysosomal degradation of protein-of-interest (POI) via cell biological and biochemical analyses.For full details on the use and execution of this protocol, please refer to Itakura et al. (2020).
format	article
author	Ayaka Tomihari Momoka Chiba Akira Matsuura Eisuke Itakura
author_facet	Ayaka Tomihari Momoka Chiba Akira Matsuura Eisuke Itakura
author_sort	Ayaka Tomihari
title	Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
title_short	Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
title_full	Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
title_fullStr	Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
title_full_unstemmed	Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
title_sort	protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/73c2926b026747bab84edd04ce386c97
work_keys_str_mv	AT ayakatomihari protocolforquantificationofthelysosomaldegradationofextracellularproteinsintomammaliancells AT momokachiba protocolforquantificationofthelysosomaldegradationofextracellularproteinsintomammaliancells AT akiramatsuura protocolforquantificationofthelysosomaldegradationofextracellularproteinsintomammaliancells AT eisukeitakura protocolforquantificationofthelysosomaldegradationofextracellularproteinsintomammaliancells
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Protocol for quantification of the lysosomal degradation of extracellular proteins into mammalian cells

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