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Protein engineering of Drosophila alcohol dehydrogenase The hydroxyl group of Tyr 152 is involved in the active site of the enzyme
Author(s) -
Gonzålez-Duarte Ricard Albalat,
Atrian Silvia
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)81282-q
Subject(s) - alcohol dehydrogenase , enzyme , tyrosine , biochemistry , active site , phenylalanine , chemistry , residue (chemistry) , alcohol , dehydrogenase , stereochemistry , amino acid
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 ban 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.