title: Over-expression of the yeast ATP synthase subunit D in Escherichia coli: use of polyclonal antibodies directed against recombinant subunit D.
abstract: The yeast ATP synthase subunit d was over-expressed in E. coli and formed inclusion bodies. It was purified by solubilization in urea and slow removal of the urea by stepwise dialysis in the presence of a non-ionic detergent. The resulting soluble subunit d was used to prepare polyclonal antibodies. Blots of yeast mitochondrial proteins were probed with these antibodies. The strain disrupted in ATP4 gene encoding the subunit 4 displayed only 8% of the wild type subunit d. Antibodies against subunit d did not inhibit the wild type ATPase activity.
authors.affiliation: Institut de Biochimie Cellulaire du CNRS, Université de Bordeaux II, France.
authors.forename: N
authors.initials: N
authors.lastname: Norais
authors.forename: J
authors.initials: J
authors.lastname: Vaillier
authors.forename: J
authors.initials: J
authors.lastname: Velours
mesh_terms: D000908:Antibodies, Fungal; D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*
chemical_list: D000908:Antibodies, Fungal; D004274:DNA, Recombinant; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D006180:Proton-Translocating ATPases