CLONING AND EXPRESSION OF AN EVOLUTIONARY CONSERVED SINGLE-DOMAIN ANGIOTENSIN-CONVERTING ENZYME FROM DROSOPHILA-MELANOGASTER
CLONING AND EXPRESSION OF AN EVOLUTIONARY CONSERVED SINGLE-DOMAIN ANGIOTENSIN-CONVERTING ENZYME FROM DROSOPHILA-MELANOGASTER
复制标题
DOI：
10.1074/jbc.270.23.13613
复制
发表时间：
1995-06-09
影响因子：
4.8
通讯作者：
ISAAC, RE
中科院分区：
文献类型：
--
作者：
CORNEL, MJ;WILLIAMS, TA;ISAAC, RE
哺乳动物体细胞血管紧张素转换酶（EC 3.4.15.1，ACE）由两个高度同源的（N-和C-）结构域组成，所述结构域由重复基因编码。我们在果蝇胚胎中发现了一种表观单结构域（67 kDa）的昆虫血管紧张素转换酶（AnCE），它将血管紧张素I转化为血管紧张素II（K-m，365 μ M），从缓激肽的C末端去除Phe-Arg（K-m，22 μ M），并被ACE抑制剂captopril（IC 50 = 1.1 × 10（-9）M）和trandolaprilat（IC 50 = 1.6 × 10（-8）M）抑制。我们还报告了果蝇AnCE cDNA的克隆和表达，该cDNA编码一个单域615个氨基酸的蛋白质，具有预测的17个氨基酸的信号肽和与哺乳动物体细胞ACE的N-和C-结构域具有高水平同源性的区域，特别是在活性位点共有序列周围。北方分析在果蝇胚胎中鉴定出单一的2.1-脱氢酶mRNA，而果蝇基因组DNA的南方分析表明昆虫基因没有复制。当在COS-7细胞中表达时，AnCE蛋白是一种分泌酶，可将血管紧张素I转化为血管紧张素II，并被卡托普利（IC 50 = 5.6 x 10（-9）M）和群多普利拉（IC 50 = 2 x 10（-8）M）抑制。这些结果的进化意义进行了讨论。
Mammalian somatic angiotensin converting enzyme (EC 3.4.15.1, ACE) consists of two highly homologous (N- and C-) domains encoded by a duplicated gene. We have identified an apparent single-domain (67 kDa) insect angiotensin converting enzyme (AnCE) in embryos of Drosophila melanogaster which converts angiotensin I to angiotensin II (K-m, 365 mu M), removes Phe-Arg from the C terminus of bradykinin (K-m, 22 mu M), and is inhibited by ACE inhibitors, captopril (IC50 = 1.1 x 10(-9) M) and trandolaprilat (IC50 = 1.6 x 10(-8) M). We also report the cloning and expression of a Drosophila AnCE cDNA which codes for a single-domain 615-amino acid protein with a predicted 17-amino acid signal peptide and regions with high levels of homology to both the N- and C-domains of mammalian somatic ACE, especially around the active site consensus sequence. Northern analysis identified a single 2.1-kilobase mRNA in Drosophila embryos, and Southern analysis of Drosophila genomic DNA indicates that the insect gene is not duplicated. When expressed in COS-7 cells, the AnCE protein is a secreted enzyme, which converts angiotensin I to angiotensin II and is inhibited by captopril (IC50 = 5.6 x 10(-9) M) and trandolaprilat (IC50 = 2 x 10(-8) M). The evolutionary significance of these results is discussed.