Nabi A H M Nurun, Kageshima Ayano, Uddin Mohammad Nasir, Nakagawa Tsutomu, Park Enoch Y, Suzuki Fumiaki
Graduate School of Agricultural Science, Gifu University, Gifu 501-1193, Japan.
Int J Mol Med. 2006 Sep;18(3):483-8.
The rat recombinant renin/prorenin receptor (AB188298 in DDJB), which conjugated with FLAG epitope in its N-terminus, was expressed in a baculovirus expression system. The recombinant receptor, prepared from the cytoplasmic fraction of the insect cells, was identified by Western blotting using anti-FLAG antibody. Prorenin as well as renin bound to the receptor with different binding affinities. Their Kd values were estimated at 8.0 and 20 nM, respectively. The amounts of prorenin and renin bound to the immobilized receptors were 1.0 and 0.2 pmole, respectively. The prorenin bound to the receptor had renin activity and the renin kept the activity at similar level to that before the binding. The Km of their complexes was the same at 3.3 microM when sheep angiotensinogen was used as the substrate. Their Vmax values were 5.5 and 10 nM.h(-1), respectively. The molecular activities of prorenin and renin bound to the receptor were 1.1 and 10 h(-1), respectively. From these findings, rat prorenin as well as renin was indicated to bind to the recombinant receptor and express the enzymatic activity in vitro.
大鼠重组肾素/前肾素受体（DDJB中的AB188298），其N端与FLAG表位偶联，在杆状病毒表达系统中表达。从昆虫细胞胞质部分制备的重组受体，用抗FLAG抗体通过蛋白质印迹法进行鉴定。前肾素以及肾素以不同的结合亲和力与该受体结合。它们的解离常数（Kd）值分别估计为8.0和20 nM。与固定化受体结合的前肾素和肾素的量分别为1.0和0.2皮摩尔。与受体结合的前肾素具有肾素活性，且肾素在结合后保持与结合前相似水平的活性。当以绵羊血管紧张素原为底物时，它们复合物的米氏常数（Km）相同，均为3.3 microM。它们的最大反应速度（Vmax）值分别为5.5和10 nM·h⁻¹。与受体结合的前肾素和肾素的分子活性分别为1.1和10 h⁻¹。从这些发现表明，大鼠前肾素以及肾素在体外均能与重组受体结合并表达酶活性。
