Campbell D J, Valentijn A J, Condron R
St Vincent's Institute of Medical Research, Fitzroy, Australia.
J Hypertens. 1991 Jan;9(1):29-33. doi: 10.1097/00004872-199101000-00005.
Rat kidney renin was purified 29,000-fold using a nine-step procedure with a 4% recovery. N-terminal sequence analysis of the non-reduced renin revealed two sequences, indicating a two-chain structure. When compared with the amino acid sequence deduced from the rat preprorenin cDNA sequence, the N-termini of the A and B chains were residues 72 and 355, respectively, of preprorenin. The two-chain structure was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Rat kidney renin migrated as two broad bands of 35,000-36,000 and 37,000-38,000 Da under non-reducing conditions. Under reducing conditions, the size of both bands decreased by approximately 3000 Da and a band migrated at the buffer front (approximately 5000 Da). The site of cleavage between the two chains of rat renin is analogous to that of mouse submandibular gland renin. However, processing of the prosequence of rat prorenin differs from that of mouse and human prorenin, indicating that the mechanism of activation of prorenin is species-specific.
使用九步程序将大鼠肾脏肾素纯化了29,000倍，回收率为4%。对未还原的肾素进行N端序列分析，发现了两个序列，表明其为双链结构。与从大鼠前肾素cDNA序列推导的氨基酸序列相比，A链和B链的N端分别是前肾素的第72位和第355位残基。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳证实了双链结构。在非还原条件下，大鼠肾脏肾素迁移为两条宽条带，分子量分别为35,000 - 36,000 Da和37,000 - 38,000 Da。在还原条件下，两条条带的大小均减小了约3000 Da，并且有一条条带迁移至缓冲液前沿（约5000 Da）。大鼠肾素两条链之间的切割位点与小鼠颌下腺肾素的类似。然而，大鼠前肾素前序列的加工与小鼠和人前肾素不同，这表明前肾素的激活机制具有物种特异性。
