A Novel Assay for Measurement of Membrane‐Protein Surface Expression using a β‐lactamase Reporter | Zendy

Lam Vincent M. | Zendy; Beerepoot Pieter | Zendy; Angers Stephane | Zendy; Salahpour Ali | Zendy

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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.

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