title: A fast and simple method to eliminate Cpn60 from functional recombinant proteins produced by E. coli Arctic Express.
abstract: A frequent problem of recombinant protein production is their insolubility. To address this issue, engineered Escherichiacoli strains like Arctic Express that produce an exogenous chaperone facilitating protein folding, have been designed. A drawback is the frequent contamination of the protein by chaperones. A simple method, using urea at a sub-denaturing concentration, allows unbinding of Cpn60 from expressed protein. This method was successfully used to purify 2 proteins, an enzyme and a viral protein. The enzyme was fully active. The nature of interaction forces between enzyme and Cpn60 was investigated. The method is likely applicable to purify other proteins.
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
authors: Lorène
authors: L
authors: Belval
authors: UMR 7177, Institut de Chimie, F-67000 Strasbourg, France.
authors: Arnaud
authors: A
authors: Marquette
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
authors: Pere
authors: P
authors: Mestre
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
authors: Marie-Christine
authors: MC
authors: Piron
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
authors: Gérard
authors: G
authors: Demangeat
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
authors: Didier
authors: D
authors: Merdinoglu
authors: INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France. Electronic address: jfchich@colmar.inra.fr.
authors: Jean-François
authors: JF
authors: Chich
mesh_terms: D001671:Biochemistry / Q000379:methods*; D018834:Chaperonin 60 / Q000378:metabolism*; D000067493:Dynamic Light Scattering; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005818:Genetic Engineering; D007700:Kinetics; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D018834:Chaperonin 60; D011994:Recombinant Proteins
keywords: Arctic Express; Cpn60; IMAC; Recombinant protein