Studying purinoceptor cell-surface expression by protein biotinylation

Young, Mark T. ORCID: https://orcid.org/0000-0002-9615-9002 2020. Studying purinoceptor cell-surface expression by protein biotinylation. Pelegrin, P., ed. Purinergic Signaling: Methods and Protocols, Vol. 2041. Methods in Molecular Biology, Springer, pp. 137-146. (10.1007/978-1-4939-9717-6_9)

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Abstract

Covalent labeling of protein with biotin (biotinylation) is a versatile technique which enables the capture and analysis of labeled protein with streptavidin-coated beads. It is particularly useful for detecting and quantifying the cell-surface expression of membrane proteins and widely used to analyze protein trafficking and the effect of mutations (particularly those which render the protein nonfunctional) on cell-surface expression. Here I describe the procedure for biotinylation and capture of cell-surface rat P2X2 receptors expressed in mammalian cells, and outline the steps in data analysis required to measure the proportion of cell-surface expressed protein of single point mutants relative to a wild-type control.

Item Type:	Book Section
Book Type:	Scholarly Edition
Date Type:	Publication
Status:	Published
Schools:	Schools > Biosciences
Publisher:	Springer
ISBN:	9781493997169
ISSN:	1064-3745
Last Modified:	26 Oct 2022 08:05
URI:	https://orca.cardiff.ac.uk/id/eprint/126547

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