title: Over-expression and refolding of beta-subunit from the chloroplast ATP synthase.
abstract: We established a bacterial system for high-level over-expression of the spinach chloroplast atpB gene which encodes the ATP synthase beta subunit. Upon induction, atpB was expressed as at least 50% to 70% of total cell protein. Although the over-expressed beta polypeptide formed insoluble inclusion bodies, more than fifty percent of it was restored to a functional form by solubilizing the inclusion bodies with 4 M urea and slowly removing the urea by stepwise dialysis. The resulting beta subunit exhibited specific and selective nucleotide binding properties identical to those of the native beta subunit.
authors.affiliation: Department of Biochemistry, University of Kansas, Lawrence 66045.
authors.forename: Z
authors.initials: Z
authors.lastname: Chen
authors.forename: I
authors.initials: I
authors.lastname: Wu
authors.forename: M L
authors.initials: ML
authors.lastname: Richter
authors.forename: P
authors.initials: P
authors.lastname: Gegenheimer
mesh_terms: D001483:Base Sequence; D002736:Chloroplasts / Q000201:enzymology*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology; D011485:Protein Binding; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry*
chemical_list: D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D006180:Proton-Translocating ATPases