title: Protein engineering of Drosophila alcohol dehydrogenase. The hydroxyl group of Tyr152 is involved in the active site of the enzyme.
abstract: Drosophila alcohol dehydrogenase is the most studied member of the family of short-chain alcohol dehydrogenases, although its tridimensional structure still remains unknown. We have engineered a Drosophila alcohol dehydrogenase in which tyrosine-152, an invariant residue in all members of the family, has been substituted by phenylalanine. The mutated gene has been expressed in yeast and pure mutant enzyme has been prepared by a one-step FPLC chromatographic procedure. Drosophila alcohol dehydrogenase-phenylalanine-152 shows no enzymatic activity. This result suggests not only that tyrosine-152 could constitute an essential building block of the active site but also that its hydroxyl group is directly involved in the redox reaction catalyzed by the enzyme.
authors.affiliation: Department of Genètica, Facultat de Biologia, Universitat de Barcelona, Spain.
authors.forename: R
authors.initials: R
authors.lastname: Albalat
authors.lastname: González-Duarte
authors.forename: S
authors.initials: S
authors.lastname: Atrian
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA; D004331:Drosophila melanogaster; D004926:Escherichia coli; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D014443:Tyrosine / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D014443:Tyrosine; D004247:DNA; D000426:Alcohol Dehydrogenase