A Rapid Method for Refolding Cell Surface Receptors and Ligands

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Date

2016-05-24

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Publisher

Nature Publishing Group

Abstract

Production of membrane-associated cell surface receptors and their ligands is often a cumbersome, expensive, and time-consuming process that limits detailed structural and functional characterization of this important class of proteins. Here we report a rapid method for refolding inclusion-body-based, recombinant cell surface receptors and ligands in one day, a speed equivalent to that of soluble protein production. This method efficiently couples modular on-column immobilized metal ion affinity purification and solid-phase protein refolding. We demonstrated the general utility of this method for producing multiple functionally active immunoreceptors, ligands, and viral decoys, including challenging cell surface proteins that cannot be produced using typical dialysis- or dilution-based refolding approaches.

Description

Keywords

inclusion-body proteins, crystal-structure, recombinant proteins, escherichia-coli, human nkg2d, cytomegalovirus, mica, purification, expression, prediction

Citation