YAKHAK HOEJI (약학회지)

The Pharmaceutical Society of Korea (대한약학회)
권호 표지

The Characteristics of I269S and I224S Double Mutant Horse Liver Alcohol Dehydrogenase

I269S와 I224S 이중변이 알코올 탈수소효소의 특성

  • Ryu, Ji-Won (College of Pharmacy, Ewha Womans University) ;
  • Lee, Kang-Man (College of Pharmacy, Ewha Womans University)
  • 류지원 (이화여자대학교 약학대학) ;
  • 이강만 (이화여자대학교 약학대학)
  • Published : 1997.12.01
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Abstract

Ile-224 in I269S mutant horse liver alcohol dehydrogenase isoenzyme S (HLADH-S) was mutated to serine by site-directed mutagenesis in order to study the role of the residue in c oenzyme binding to the enzyme. The specific activity of the I269S and I224S mutant enzyme to ethanol was increased 6-fold and all Michaelis constants($K_a,\;K_b,\;K_p,\;and\;K_q$,/TEX>) were larger than those for the wild-type and I269S enzyme. The substitution decreased the afffinity to coenzymes and increased the specific activity of the enzyme. The mutant enzyme showed the highest catalytic efficiency for octanol among the primary alcohols. But it didn`t have activities on retinoids and 5${\beta}$-cholanic acid-3-one. From these results, it was confirmed that the hydrophobic interaction of Ile-224 residue with coenzyme was related to coenzyme affinity in ADH reaction. The substitution also affected the substrate affinities to the enzyme.

Keywords

References

  1. Biochemistry v.2 Product inhibition studies on yeast and liver alcohol dehydrogenase Wratten, C. C.;Cleland, W. W.
  2. J. Biochem. v.171 Specificities and configurations of ternary complexes of yeast and liver alcohol dehydrogenase Dickinson, F. M.;Dickenson, C. J.
  3. J. Biol. Chem. v.257 Binding of substrate in a ternary complex of horse liver alcohol dehydrogenase Eklund, H.;Plapp, B. V.;Samama, J. P.;Branden, C. I.
  4. J. Mol. Biol. v.102 Three-dimensional structure of horse liver alcohol dehydrogenase at 2-4 A resolution Eklund, H.;Nordstrom, B.;Zeppezauer, E.;Soderlund, G.;Ohlsson, I.;Boiwe, T.;Soderberg, B. O.;Tapia, O.;Branden, C. I.;Akeson, A.
  5. J. Biochem. v.254 Structural difference between apo and holoenzyme of horse liver alcohol dehydrogenase Eklund, H.;Branden, C. I.
  6. J. Mol. Biol. v.146 Structure of triclinic ternary complex of horse liver alcohol dehydrogenase at 2.9 A resolution Eklund, H.;Samama, J. P.;Wallen, L.;Branden, C. I.;Akeson, A.;Jones, T. A.
  7. Biochemistry v.33 Structures of horse liver alcohol dehydrogenase complexed with NAD^{+}$ and substituted benzyl alcohols Ramaswamy, S.;Eklund, H.;Plapp, B. V.
  8. J. Biol. Chem. v.266 Isoenzymes of horse liver alcohol dehydrogenase active on ethanol and steroids. cDNA cloning, expression, and comparison of active sites Park, D. H.;Plapp, B. V.
  9. J. Biol. Chem. v.270 no.9 Genomic structure and expression of the ADH 7 gene encoding human class Ⅳ alcohol dehydrogenase, the form most efficient for retinol metabolism in vitro Mirna, Z.;Mario, H. F.;Gregg, D.
  10. Anal. Biochem. v.214 Spectrophotometric assay for α-ketoaldehydes using horse liver alcohol dehydrogenase Yang, C. F.;Brush, E. J. A.
  11. Arch. Biochem. Biophysics v.225 no.2 Separation and partial characterization of multiple forms of rat liver alcohol dehydrogenase Mezey, E.;Potter, J. J.
  12. J. Biol. Chem. v.258 no.5 Physiological function and kinetic mechanism of human liver alcohol dehydrogenase as 5β-cholestane-3α, 7α, 12α, 26α-tetrol dehydrogenase Akihiko, O.;Kyuichiro, O.
  13. FEBS Letters v.257 no.1 Evidence for the identity of glutathione-dependent formaldehyde dehydrogenase and class Ⅲ alcohol dehydrogenase Koivusalo, M. K.;Baumann, M. B.;Uotila, L.
  14. Biochemistry v.33 Residues specific for class III alcohol dehydrogenase site-directed mutagenesis of the human enzyme Estonius, M.;Hoog, J. O.;Danielsson, O.;Jornvall, H.
  15. Eur. J. Biochem. v.162 Characterization of three isoenzymes of rat alcohol dehydrogenase: tissue distribution and physical and enzymatic properties Julia, P.;Farres, J.;Pares, X.
  16. Method. Enzymol. v.189 Isoenzymes of alcohol dehydrogenase in retinoid metabolism Pares, X.;Julia, P.
  17. Arch. Biochem. Biophysics. v.307 no.1 Physiological substrates for rat alcohol dehydrogenase classes: aldehydes of lipid peroxidation, ω-hydroxy fatty acids and retinoids Boleda, M. D.;Saubi, N.;Farres, J.;Pares, X.
  18. J. Biol. Chem. v.270 no.8 Expression and kinetic characterization of recombinant human stomach alcohol dehydrogenase: active-site amino acid sequence explains substrate specificity compared with liver isozymes Kedisshvili, N. Y.;Bosron, W. F.;Stone, C. L.;Hurley, T. D.;Peggs, C. F.;Thomasson, H. R.;Popov, K. M.;Carr, L. G.;Edenberg, H. J.;Li, T. K.
  19. Arch. Pharm. Res. v.20 no.2 I269S mutation in horse liver alcohol dehydrogenase S isoenzyme and its reactivity for steroids and retinoids Ryu, J. W.;Lee, K. M.
  20. Anal. Biochem. v.200 Site-directed mutagenesis of virtually any plasmid by eliminating a unique site Deng, W. P.;Nickoloff, J. A.
  21. Proc. Natl. Acad. Sci. USA v.74 DNA sequecing with chain-terminating inhibitors Sanger, F.;Niklen, S.;Coulson, A. R.
  22. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  23. J. Biol. Chem. v.245 Enhancement of the activity of horse liver alcohol dehydrogenase by modification of amino groups at the active sites Plapp, B. V.
  24. PH. D. THESIS, The University of Iowa Structure and function of isoenzyme of horse liver alcohol dehydrogenase Park, D. H.
  25. J. Biol. Chem. v.267 Interconversion of E and S isoenzyme of horse liver alcohol dehydrogenase Park, D. H.;Plapp, B. P.
  26. Biochemistry v.16 Kinetics of native and activated isozymes of horse liver alcohol dehydrogenase Dworschack, R. T.;Plapp, B. V.
  27. Biochemistry v.19 Kinetic mechanism of horse liver alcohol dehydrogenase SS Ryzewski, C. N.;an Pietruszko, R.
  28. Biochemistry v.29 Substitution of arginine for histidine-47 in the coenzyme binding site of yeast alcohol dehydrogenase Ⅰ Gould, R. M.;Plapp, B. V.