Artículo
Dynamic Distribution of ASIC1a Channels and Other Proteins within Cells Detected through Fractionation
Salinas Castellanos, Libia Catalina; Gatto, Rodolfo Gabriel; Menchón, Silvia Adriana
; Blaustein, Matías; Uchitel, Osvaldo Daniel
; Weissmann, Carina
Fecha de publicación:
03/2022
Editorial:
MDPI
Revista:
Membranes
ISSN:
2077-0375
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Proteins in eukaryotic cells reside in different cell compartments. Many studies require the specific localization of proteins and the detection of any dynamic changes in intracellular protein distribution. There are several methods available for this purpose that rely on the fractionation of the different cell compartments. Fractionation protocols have evolved since the first use of a centrifuge to isolate organelles. In this study, we described a simple method that involves the use of a tabletop centrifuge and different detergents to obtain cell fractions enriched in cytosolic (Cyt), plasma membrane (PM), membranous organelle (MO), and nuclear (Nu) proteins and identify the proteins in each fraction. This method serves to identify transmembrane proteins such as channel subunits as well as PM-embedded or weakly associated proteins. This protocol uses a minute amount of cell material and typical equipment present in laboratories, and it takes approximately 3 h. The process was validated using endogenous and exogenous proteins expressed in the HEK293T cell line that were targeted to each compartment. Using a specific stimulus as a trigger, we showed and quantified the shuttling of a protein channel (ASIC1a, acid sensing ion channel) from the MO fraction to the PM fraction and the shuttling of a kinase from a cytosolic location to a nuclear location.
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Articulos(IFEG)
Articulos de INST.DE FISICA ENRIQUE GAVIOLA
Articulos de INST.DE FISICA ENRIQUE GAVIOLA
Articulos(IFIBYNE)
Articulos de INST.DE FISIOL., BIOL.MOLECULAR Y NEUROCIENCIAS
Articulos de INST.DE FISIOL., BIOL.MOLECULAR Y NEUROCIENCIAS
Articulos(OCA CIUDAD UNIVERSITARIA)
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA CIUDAD UNIVERSITARIA
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA CIUDAD UNIVERSITARIA
Citación
Salinas Castellanos, Libia Catalina; Gatto, Rodolfo Gabriel; Menchón, Silvia Adriana; Blaustein, Matías; Uchitel, Osvaldo Daniel; et al.; Dynamic Distribution of ASIC1a Channels and Other Proteins within Cells Detected through Fractionation; MDPI; Membranes; 12; 4; 3-2022; 1-12
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