Albert V R, Allen J M, Joh T H
J Biol Chem. 1987 Jul 5;262(19):9404-11.
We have sought to determine whether aromatic L-amino acid decarboxylase which functions as a neurotransmitter biosynthetic enzyme in neuronal cells can be distinguished from an enzyme with similar activity found in peripheral tissues where no neurotransmitters are synthesized. Aromatic L-amino acid decarboxylase was purified to electrophoretic homogeneity from bovine adrenal medulla, and highly specific antibodies were produced. In addition, a DNA clone complementary to aromatic L-amino acid decarboxylase mRNA was isolated by immunological screening of a lambda gt11 cDNA expression library. We have used these antibodies and cDNA probes for biochemical, immunochemical, and molecular analyses. A single form of aromatic L-amino acid decarboxylase is detected in rat and bovine tissue. Specifically, aromatic L-amino acid decarboxylase protein is biochemically and immunochemically indistinguishable in brain, liver, kidney, and adrenal medulla. Hybridization to aromatic L-amino acid decarboxylase cDNA identifies a single mRNA species of 2.3 kilobase pairs in rat tissue. Furthermore, Southern blot analysis reveals that a single gene codes for aromatic L-amino acid decarboxylase.
我们试图确定在神经元细胞中作为神经递质生物合成酶发挥作用的芳香族L-氨基酸脱羧酶，是否能与在不合成神经递质的外周组织中发现的具有类似活性的酶区分开来。从牛肾上腺髓质中将芳香族L-氨基酸脱羧酶纯化至电泳纯，并制备了高度特异性的抗体。此外，通过对λgt11 cDNA表达文库进行免疫筛选，分离出了与芳香族L-氨基酸脱羧酶mRNA互补的DNA克隆。我们已将这些抗体和cDNA探针用于生化、免疫化学和分子分析。在大鼠和牛组织中检测到单一形式的芳香族L-氨基酸脱羧酶。具体而言，芳香族L-氨基酸脱羧酶蛋白在脑、肝、肾和肾上腺髓质中的生化性质和免疫化学性质无法区分。与芳香族L-氨基酸脱羧酶cDNA杂交可鉴定出大鼠组织中一种2.3千碱基对的单一mRNA种类。此外，Southern印迹分析表明，单一基因编码芳香族L-氨基酸脱羧酶。
