An attempt to transform class characteristics within the alcohol dehydrogenase family
| Autor: | Jesper J. Hedberg, Jan-Olov Höög, Patrik Strömberg |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Recombinant protein
Immunoblotting Biophysics Pyruvate dehydrogenase phosphatase Arginine Biochemistry Formaldehyde dehydrogenase activity chemistry.chemical_compound Leucine Structural Biology Serine Genetics Humans Molecular Biology Formaldehyde dehydrogenase Alcohol dehydrogenase Binding Sites Ethanol biology Lysine Enzyme kinetics Lauric Acids ADH1B Cell Biology Aldehyde Oxidoreductases Glutathione Recombinant Proteins Kinetics chemistry Mutagenesis Mutation biology.protein Oxoglutarate dehydrogenase complex Branched-chain alpha-keto acid dehydrogenase complex |
| Zdroj: | FEBS Letters. 436:67-70 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/s0014-5793(98)01100-4 |
| Popis: | Human class I alcohol dehydrogenase was mutated at positions 57 and 115, exchanging for Asp and Arg respectively, in an attempt to introduce glutathione-dependent formaldehyde dehydrogenase characteristics. In addition, class III alcohol dehydrogenase, identical to glutathione-dependent formaldehyde dehydrogenase, was mutated at position 115, introducing Ser or Lys. The attempted class transformation was partly successful considering a higher affinity for 12-hydroxydodecanoate and a lower affinity for ethanol that was monitored for the class I mutant. However, the class I mutant displayed neither glutathione-dependent formaldehyde dehydrogenase activity nor fatty acid activation of alcohol oxidation. Interestingly, both class III mutants showed reduced activities for S -hydroxymethylglutathione and 12-hydroxydodecanoate through increased K m values. Overall results show that it is not possible, by single point mutations, to completely transform enzyme characteristics between these two classes of alcohol dehydrogenase. |
| Databáze: | OpenAIRE |
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