Cloning of a rat cDNA encoding retinal dehydrogenase isozyme type I and its expression in E. coli.
Cloning of a rat cDNA encoding retinal dehydrogenase isozyme type I and its expression in E. coli.
复制标题
编码视网膜脱氢酶同工酶 I 型的大鼠 cDNA 的克隆及其在大肠杆菌中的表达。
DOI：
10.1016/s0378-1119(97)00054-1
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发表时间：
1997
期刊：
影响因子：
3.5
通讯作者：
Napoli,JL
中科院分区：
文献类型：
--
作者：
Penzes,P;Wang,X;Sperkova,Z;Napoli,JL
从纯化的大鼠肝细胞溶质视网膜脱氢酶 P1 中测序的肽 [Posch, K.C.、Burns, R.D. 和 Napoli, J.L., 1992。从视网膜生物合成全反式视黄酸：纯化的细胞溶质脱氢酶识别与细胞视黄醇结合蛋白（I 型）结合的视网膜作为底物。 J.Biol。化学。 267, 19676–19682] 用于设计用于克隆其 cDNA 的寡核苷酸。 P1 的推导氨基酸序列现在被命名为 I 型视网膜脱氢酶或 RalDH(I)，与小鼠 AHD-2 和大鼠肾乙醛脱氢酶非常相似，但与大鼠苯巴比妥诱导乙醛脱氢酶 (PIADH) 不同，PIADH 是小鼠 AHD-2 的假定大鼠肝脏同源物。大鼠肾脏(100%)和肺(88%)的RalDH(I) mRNA水平相对较高，肝脏(34%)和大脑(22%)的水平中等，睾丸(8%)的水平较低。类维生素A状态对不同组织中RalDH(I) mRNA水平的影响不同。大肠杆菌表达的 RalDH(I) 对视网膜表现出变构动力学，Hill 系数为 1.7，K0.5 值为 1.4 μM，Vmax 为 52 nmol min−1mg−1 蛋白。这些数据建立了 P1 和 RalDH(I) 的共特异性，表明类维生素A状态以组织依赖性方式影响其 mRNA 的表达，并说明具有广泛同源性的乙醛脱氢酶同工酶可以参与不同的代谢途径，例如 RalDH 与 PIADH。
Peptides sequenced from the purified rat liver cytosolic retinal dehydrogenase P1 [Posch, K.C., Burns, R.D. and Napoli, J.L., 1992. Biosynthesis of all-trans-retinoic acid from retinal: recognition of retinal bound to cellular retinol-binding protein (type I) as substrate by a purified cytosolic dehydrogenase. J. Biol. Chem. 267, 19676–19682] were used to design oligonucleotides for cloning its cDNA. The deduced amino acid sequence of P1, now designated retinal dehydrogenase type I or RalDH(I), has close similarity with mouse AHD-2 and rat kidney aldehyde dehydrogenase, but is distinct from rat phenobarbital-inducible aldehyde dehydrogenase (PIADH), the presumed rat liver homolog of mouse AHD-2. Rat kidney (100%) and lung (88%) show relatively high mRNA levels of RalDH(I), liver (34%) and brain (22%) have moderate levels, and testis (8%) has low levels. Retinoid status affects RalDH(I) mRNA levels differently in different tissues. E. coli-expressed RalDH(I) exhibits allosteric kinetics for retinal with a Hill coefficient of 1.7, a K0.5value of 1.4 μM and a Vmaxof 52 nmol min−1mg−1protein. These data establish the cospecificity of P1 and RalDH(I), show that retinoid status affects expression of its mRNA in a tissue-dependent manner, and illustrate that aldehyde dehydrogenase isozymes with extensive homology can participate in different metabolic paths, e.g., RalDH vs. PIADH.