Carsia R V, McIlroy P J, Kowalski K I, Tilly J L
Department of Cell Biology, University of Medicine and Dentistry of New Jersey-School of Osteopathic Medicine, Stratford 08084.
Biochem Biophys Res Commun. 1993 Mar 31;191(3):1073-80. doi: 10.1006/bbrc.1993.1326.
A turkey adrenocortical cell AII receptor cDNA fragment (714 bp) was isolated by RT-PCR using oligonucleotide primers based on rat aortic smooth muscle (RASM) and bovine adrenal type-1 (AT1) receptor cDNA coding sequences as primers. Sequence analysis indicated 73% nucleotide identity and 78% amino acid identity to the RASM AT1 receptor. Notable differences were 1) two additional Cys at positions 92 and 99 (first extracellular loop), 2) deletion of amino acid 168, formation of a triplet Asn sequence (Asn 186, 187, 188) and substitution of Arg192 with Pro (second extracellular loop) and 3) two additional potential protein kinase C phosphorylation sites, Thr221 and Thr233 (third intracellular loop). Southern blot analysis indicated that the receptor is a product of a single-copy gene. Northern blot analysis indicated at least three mRNA transcripts (7.5, 3.5 and 2.0 kb) expressed predominantly in the adrenal gland.
利用基于大鼠主动脉平滑肌(RASM)和牛肾上腺1型(AT1)受体cDNA编码序列的寡核苷酸引物,通过逆转录聚合酶链反应(RT-PCR)分离出一段火鸡肾上腺皮质细胞AII受体cDNA片段(714 bp)。序列分析表明,该片段与RASM AT1受体的核苷酸同一性为73%,氨基酸同一性为78%。显著差异包括:1)在第92和99位(第一个细胞外环)有两个额外的半胱氨酸;2)氨基酸168缺失,形成一个三联体天冬酰胺序列(天冬酰胺186、187、188),并且第192位的精氨酸被脯氨酸取代(第二个细胞外环);3)在第221和233位(第三个细胞内环)有两个额外的潜在蛋白激酶C磷酸化位点。Southern印迹分析表明该受体是单拷贝基因的产物。Northern印迹分析表明至少有三种mRNA转录本(7.5、3.5和2.0 kb),主要在肾上腺中表达。