Labrecque J, Bhat P V, Lacroix A
Department of Medicine, University of Montreal, Que., Canada.
Biochem Cell Biol. 1993 Jan-Feb;71(1-2):85-9. doi: 10.1139/o93-013.
A NAD-dependent aldehyde dehydrogenase (EC 1.2.1.3) which catalyzes the oxidation of retinal to retinoic acid was purified to homogeneity from rat kidney by using Affi-Gel blue affinity chromatography and chromatofocusing, followed by Mono-Q anion-exchange chromatography. The apparent molecular weight of the native enzyme determined by size-exclusion fast protein liquid chromatography was 140,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave a subunit molecular weight of 53,000. The isoelectric point as measured by chromatofocusing was 8.5. The enzyme also catalyzed the oxidation of acetaldehyde, but showed much lower Km value for the retinal substrate. We suggest that aldehyde dehydrogenase found in the kidney may be a specific retinal dehydrogenase, involved in vitamin A metabolism.
通过使用Affi-Gel蓝亲和色谱法和色谱聚焦，随后进行Mono-Q阴离子交换色谱法，从大鼠肾脏中纯化出一种NAD依赖性醛脱氢酶（EC 1.2.1.3），该酶催化视黄醛氧化为视黄酸。通过尺寸排阻快速蛋白质液相色谱法测定的天然酶的表观分子量为140,000。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳给出的亚基分子量为53,000。通过色谱聚焦测量的等电点为8.5。该酶还催化乙醛的氧化，但对视黄醛底物的Km值要低得多。我们认为在肾脏中发现的醛脱氢酶可能是一种特定的视黄醛脱氢酶，参与维生素A代谢。
