Morpeth F F, Dickinson F M
Biochem J. 1980 Nov 1;191(2):619-26. doi: 10.1042/bj1910619.
Aldehyde reductase was purified from pig kidney cortex to homogeneity by a new procedure. The molecular weight of the enzyme was estimated by sedimentation equilibrium to be 43 700 and by gel electrophoresis in the presence of sodium dodecyl sulphate to be 41 700. The enzyme is clearly a monomer. The enzyme preparation contained no significant quantities of zinc, manganese or copper and had no essential histidine or thiol groups. Changes in the absorption and fluorescence spectra of NADPH were observed on formation of the enzyme-NADPH complexes. Large changes in the fluorescence spectra were also observed in the presence of sodium barbitone or Warfarin. These changes were used as the basis of active-site titrations, which showed that the enzyme had one active site per molecule. The dissociation constants of NADPH and NADP+ from binary complexes with the enzyme were estimated in spectrophotometric titrations.
通过一种新方法从猪肾皮质中纯化醛还原酶至同质。通过沉降平衡估计该酶的分子量为43700,在十二烷基硫酸钠存在下通过凝胶电泳估计为41700。该酶显然是单体。酶制剂中不含大量的锌、锰或铜,也没有必需的组氨酸或巯基基团。在形成酶-NADPH复合物时观察到NADPH吸收光谱和荧光光谱的变化。在巴比妥钠或华法林存在下也观察到荧光光谱的巨大变化。这些变化被用作活性位点滴定的基础,结果表明该酶每个分子有一个活性位点。在分光光度滴定中估计了NADPH和NADP +与酶的二元复合物的解离常数。