Cell-surface Biotinylation Assay

A cell can regulate the amount of particular proteins on its cell membrane through endocytosis, following which cell surface proteins are effectively sequestered in the cytoplasm. Once within a cell, these surface proteins can be either destroyed or "recycled" back to the membrane. The cel...

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Detalles Bibliográficos
Formato: Video
Idioma:Inglés
Publicado: Cambridge, MA : MyJoVE Corp 2016.
Colección:JOVE Science Education.
Cell Biology.
Acceso en línea:Acceso a vídeo desde UNAV
Ver en Universidad de Navarra:https://innopac.unav.es/record=b42116624*spi
Descripción
Sumario:A cell can regulate the amount of particular proteins on its cell membrane through endocytosis, following which cell surface proteins are effectively sequestered in the cytoplasm. Once within a cell, these surface proteins can be either destroyed or "recycled" back to the membrane. The cell surface biotinylation assay provides researchers with a way to study these phenomena. The technique makes use of a derivative of the small molecule biotin, which can label surface proteins and then be chemically cleaved. However, if the surface protein is endocytosed, the biotin derivative will be protected from cleavage. Thus, by analyzing the uncleaved, endocytosed biotin label, scientists can assess the amounts of internalized surface proteins.In this video, we review the concepts behind the biotinylation assay, delving into the chemical structure of the biotin derivative and the mechanism of its cleavage. This is followed by a generalized protocol of the technique, and finally, a description of how researchers are currently using it to study the dynamics of different cell surface proteins.
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Descripción Física:1 recurso electrónico (552 seg.) : son., col
Formato:Forma de acceso: World Wide Web.
Público:Para estudiantes universitarios, graduados y profesionales.