A Proximity-Based Assay for Identification of Ligand and Membrane Protein Interaction in Living Cells

Hongkai Zhang; Richard A Lerner

doi:10.1007/978-1-4939-6857-2_13

A Proximity-Based Assay for Identification of Ligand and Membrane Protein Interaction in Living Cells

Methods Mol Biol. 2017:1575:215-222. doi: 10.1007/978-1-4939-6857-2_13.

Authors

Hongkai Zhang¹, Richard A Lerner²

Affiliations

¹ Department of Cell and Molecular Biology, The Scripps Research Institute, Mail Code: MB 10, 10550 N. Torrey Pines Rd, La Jolla, CA, 92037, USA.
² Department of Cell and Molecular Biology, The Scripps Research Institute, Mail Code: MB 10, 10550 N. Torrey Pines Rd, La Jolla, CA, 92037, USA. rlerner@scripps.edu.

PMID: 28255883
DOI: 10.1007/978-1-4939-6857-2_13

Abstract

There is a need for a general reporter system to study the interactions between a ligand and membrane protein in living cells. Here, we demonstrate a method that allows identification of undiscovered ligand partners of a membrane protein in its natural milieu. This assay will be of importance especially for multiple pass transmembrane proteins such as ion channels and GPCRs.

Keywords: Interaction; Lentivirus; Ligand; Membrane protein; Proximity-based assay; Reporter cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endopeptidases / genetics
Endopeptidases / metabolism
HEK293 Cells
Humans
Ligands
Protein Binding
Protein Interaction Mapping / methods*
Receptors, Platelet-Derived Growth Factor / chemistry
Receptors, Platelet-Derived Growth Factor / genetics*
Receptors, Platelet-Derived Growth Factor / metabolism*
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism

Substances

Ligands
Recombinant Fusion Proteins
Receptors, Platelet-Derived Growth Factor
Endopeptidases
TEV protease