Structural properties of long- and short-chain alcohol dehydrogenases. Contribution of NAD+ to stability.
Structural studies were undertaken on long-chain and short-chain alcohol dehydrogenases (from horse liver and Drosophila respectively). Far-u.v. c.d. measurements were used to estimate the secondary structure contents of the enzymes. For the horse liver enzyme, the results agree well with the X-ray data; for the Drosophila enzyme (for which a crystal structure is not yet available), the results are in good agreement with those obtained by applying a range of structure-prediction procedures to the amino acid sequence of this enzyme. The conformational stabilities of the two enzymes were investigated by studying the unfolding brought about by guanidinium chloride (GdnHCl) by using activity and c.d. measurements. The unfolding of the Drosophila enzyme was analysed in terms of a two-state model; the presence of the substrate NAD+ leads to considerable protection against unfolding. By contrast, the unfolding of the horse liver enzyme shows a plateau effect at intermediate concentrations of GdnHCl, indicating that a two-state model is not appropriate in this case. NAD+ affords little, if any, protection against unfolding for the horse liver enzyme.
Ribas De Pouplana
L
L
Department of Biochemistry, University of Edinburgh, Scotland, U.K.
Atrian
S
S
Gonzàlex-Duarte
R
R
Fothergill-Gilmore
L A
LA
Kelly
S M
SM
Price
N C
NC
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002942:Circular Dichroism; D004330:Drosophila / Q000201:enzymology; D004331:Drosophila melanogaster / Q000201:enzymology; D004795:Enzyme Stability; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D006736:Horses; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D009243:NAD / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013379:Substrate Specificity
D006146:Guanidines; D009243:NAD; D000426:Alcohol Dehydrogenase; D019791:Guanidine