Redesign of the coenzyme specificity of a dehydrogenase by protein engineering
- 1 January 1990
- journal article
- research article
- Published by Springer Nature in Nature
- Vol. 343 (6253) , 38-43
- https://doi.org/10.1038/343038a0
Abstract
Directed mutagenesis and molecular modelling have been used to identify a set of amino-acid side chains in glutathione reductase that confer specificity for the coenzyme NADP+. Systematic replacement of these amino acids, all of which occur in a 'fingerprint' structural motif in the NADP+-binding domain, leaves the substrate specificity unchanged but converts the enzyme into one displaying a marked preference for the coenzyme NAD+.Keywords
This publication has 36 references indexed in Scilit:
- Alteration of mouse cytochrome P450coh substrate specificity by mutation of a single amino-acid residueNature, 1989
- From analysis to synthesis: new ligand binding sites on the lactate dehydrogenase framework. Part ITrends in Biochemical Sciences, 1989
- Engineering of an intersubunit disulphide bridge in glutathione reductase from Escherichia coliFEBS Letters, 1988
- Purification and characterization of glutathione reductase encoded by a cloned and over-expressed gene in Escherichia coliBiochemical Journal, 1987
- Refined structure of glutathione reductase at 1.54 Å resolutionJournal of Molecular Biology, 1987
- Selective alteration of substrate specificity by replacement of aspartic acid-189 with lysine in the binding pocket of trypsinBiochemistry, 1987
- Reversal of substrate charge specificity by site-directed mutagenesis of aspartate aminotransferaseJournal of the American Chemical Society, 1987
- Glutathione reductase from Escherichia coli: cloning and sequence analysis of the gene and relationship to other flavoprotein disulfide oxidoreductasesBiochemistry, 1986
- THIOREDOXINAnnual Review of Biochemistry, 1985
- Three-dimensional structure of glutathione reductase at 2 Å resolutionJournal of Molecular Biology, 1981