Drosophila lebanonensis ADH: analysis of recombinant wild‐type enzyme and site‐directed mutants
- 21 March 1994
- journal article
- Published by Wiley in FEBS Letters
- Vol. 341 (2-3) , 171-176
- https://doi.org/10.1016/0014-5793(94)80451-6
Abstract
Unique amino acid substitutions occur in D. lebanonensis ADH. They are found within the putative NAD + -binding domain and affect residues that are otherwise highly conserved in all other species of the genus. To restore the consensus ainino acids, we have constructed an expression system for this enzyme in E. coli, and engineered two mutants, Ala13 Gly and Asn56 Thr. The biochemical and kinetic features of these retromutants are consistent with increased catalytic efficiency and thermal stability. Thus, results show that wild-type D. lebanonensis ADH can be improved by site-directed mutagenesis.Keywords
This publication has 24 references indexed in Scilit:
- Site-specific mutagenesis of Drosophila alcohol dehydrogenase: Evidence for involvement of tyrosine-152 and lysine-156 in catalysisBiochemistry, 1993
- Effect of site‐directed mutagenesis on conserved positions of Drosophila alcohol dehydrogenaseFEBS Letters, 1993
- Alcohol Dehydrogenases: Patterns of Protein EvolutionPublished by Springer Nature ,1993
- Short‐chain dehydrogenasesEuropean Journal of Biochemistry, 1992
- Protein engineering of Drosophila alcohol dehydrogenase The hydroxyl group of Tyr152 is involved in the active site of the enzymeFEBS Letters, 1992
- Role of aspartic acid 38 in the cofactor specificity of Drosophila alcohol dehydrogenaseEuropean Journal of Biochemistry, 1991
- Characteristics of short‐chain alcohol dehydrogenases and related enzymesEuropean Journal of Biochemistry, 1991
- Site-directed mutagenesis of glycine-14 and two "critical" cysteinyl residues in Drosophila alcohol dehydrogenaseBiochemistry, 1990
- Alcohol and polyol dehydrogenases are both divided into two protein types, and structural properties cross-relate the different enzyme activities within each type.Proceedings of the National Academy of Sciences, 1981
- Three-dimensional structure of horse liver alcohol dehydrogenase at 2.4 Å resolutionJournal of Molecular Biology, 1976