title: A Rapid Method for Refolding Cell Surface Receptors and Ligands.
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.
authors.affiliation: Department of Biochemistry, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.|Interdepartmental Microbiology Graduate Program, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhai
authors.affiliation: Department of Biochemistry, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.
authors.forename: Ling
authors.initials: L
authors.lastname: Wu
authors.affiliation: Department of Biochemistry, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Biochemistry, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Burnham
authors.affiliation: Department of Tropical Medicine, Tulane University, New Orleans, LA 70112, USA.
authors.forename: Juan C
authors.initials: JC
authors.lastname: Pizarro
authors.affiliation: Department of Biochemistry, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.|Interdepartmental Microbiology Graduate Program, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.|Center for Drug Discovery, Virginia Polytechnic Institute &State University, Blacksburg, VA 24061, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Xu
mesh_terms: D002942:Circular Dichroism; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D008958:Models, Molecular; D058849:Protein Refolding; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D008024:Ligands; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins