Analyse cinétique des rétinaldéhydes déshydrogénases recombinantes de type 3 et 4 de souris
SUMMARY         Retinal dehydrogenases (RALDHs) catalyze the dehydrogenation of retinal into retinoic acids (RA) that are required for embryogenesis and tissue differentiation. This study sought to determine the detailed kinetic properties of 2 mouse RALDHs, namely RALDH3 and 4, for retinal isomer substrates, to better define their specificities in RA isomer synthesis.          RALDH3 and 4 were expressed as His-tagged proteins and affinity-purified. RALDH3 oxidized all-trans retinal with high catalytic efficiency but did not show activity for either 9-cis or 13-cis retinal substrates. RALDH4 was inactive for all-trans retinal substrate, exhibited high activity for 9-cis retinal oxidation, and oxidized 13-cis retinal with lower catalytic efficiency.          β-ionone, a potent inhibitor of RALDH4 activity, suppressed 9-cis and 13-cis retinal oxidation competitively, but had no effect on RALDH3 activity. The p-HMB inhibited the activity for both RALDH3 and RALDH4. The divalent cation MgCl2 activated 13-cis retinal oxidation by RALDH4 by 3-fold, slightly decreased 9-cis retinal oxidation, and did not significantly influence RALDH3 activity.         These data extend the kinetic characterization of RALDH3 and 4, providing their specificities for retinal isomer substrates, which should help in determining their functions in the synthesis of RAs in specific tissues.
Retinoic acids Aldehyde dehydrogenases Retinal isomers Retinal dehydrogenases β-Ionone β-Ionone