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A Novel Assay for Measurement of Membrane‐Protein Surface Expression using a β‐lactamase Reporter
Author(s) -
Lam Vincent M.,
Beerepoot Pieter,
Angers Stephane,
Salahpour Ali
Publication year - 2013
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12073
Subject(s) - biology , microbiology and biotechnology , protein expression , membrane , reporter gene , membrane protein , surface (topology) , computational biology , biochemistry , gene expression , gene , geometry , mathematics
Synopsis The trafficking of membrane proteins is dynamic and contributes to the homeostatic control of their cell surface localization and their function in signal transduction. Here, we describe a simple, low cost, and potentially high‐throughput assay, for measurement of surface expression of proteins. The assay uses β‐lactamase enzyme (βlac) as a reporter, and its cell‐impermeable substrate, nitrocefin. The utility of this assay is demonstrated using well‐established paradigms of internalization and molecular chaperoning, applied to two GPCRs and a monoamine transporter.