A Rapid Method for Refolding Cell Surface Receptors and Ligands

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-base...

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Bibliographic Details
Published in:Scientific reports 2016-05, Vol.6 (1), p.26482-26482, Article 26482
Main Authors: Zhai, Lu, Wu, Ling, Li, Feng, Burnham, Robert S., Pizarro, Juan C., Xu, Bin
Format: Article
Language:English
Subjects:
631/1647
631/1647/2196
82/16
82/80
82/83
Cell surface
Circular Dichroism
Dialysis
Humanities and Social Sciences
Inclusion Bodies - chemistry
Inclusion Bodies - metabolism
Ligands
Models, Molecular
multidisciplinary
Protein folding
Protein purification
Protein Refolding
Receptors, Cell Surface - chemistry
Receptors, Cell Surface - isolation & purification
Recombinant Proteins - chemistry
Science
Science (multidisciplinary)
Structure-function relationships
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep26482