Biswas Kazal Boron, Nabi A H M Nurun, Arai Yoshie, Nakagawa Tsutomu, Ebihara Akio, Suzuki Fumiaki
United Graduate School of Agricultural Science, Gifu University, Gifu 501-1193, Japan.
Front Biosci (Elite Ed). 2010 Jun 1;2(4):1234-40. doi: 10.2741/e183.
In this study, binding properties of human prorenin to rat (pro)renin receptor (r(P)RR) and rat prorenin to human (pro)renin receptor (h(P)RR) were elucidated in vitro to investigate species specificity of prorenin bindings to (P)RR. Both (P)RRs were expressed in vitro based on wheat germ lysate and purified using His trap column. The association and dissociation rate constants of human and rat prorenin for the immobilized r(P)RR and h(P)RR were 6.64 x 10(6) and 1.01 x 10(6) M-1.s-1 and, 0.024 and 8.45 x 10(-3) s-1, respectively. Their KD values were 3.7 nM (3-fold higher than that of human prorenin vs h(P)RR (1.2 nM)) and 8.3 nM (1.2-fold lower than that of rat prorenin vs r(P)RR (10.2 nM)), respectively. Additionally, human and rat prorenin bound to the pre-adsorbed rat and human (P)RR, respectively, performed enzymatic activities. Their molecular activities were 4.1h-1 (almost similar (3.8h -1) to human prorenin vs h(P)RR) and 0.98 h-1 (approximately 2-fold lower than rat prorenin vs r(P)RR), respectively. Thus, these results suggest the species specificity for bindings of prorenin to (P)RR, which could be useful in elucidating the biochemical properties of prorenin binding to the receptor.
在本研究中，体外阐明了人肾素原与大鼠（原）肾素受体（r(P)RR）以及大鼠肾素原与人（原）肾素受体（h(P)RR）的结合特性，以研究肾素原与（P)RR结合的物种特异性。两种（P)RR均基于麦胚裂解物在体外表达，并使用His trap柱进行纯化。人肾素和大鼠肾素与固定化r(P)RR和h(P)RR的缔合和解离速率常数分别为6.64×10⁶和1.01×10⁶ M⁻¹·s⁻¹以及0.024和8.45×10⁻³ s⁻¹。它们的KD值分别为3.7 nM（比人肾素原与人（P)RR的结合（1.2 nM）高3倍）和8.3 nM（比大鼠肾素原与r(P)RR的结合（10.2 nM）低1.2倍）。此外，人肾素和大鼠肾素分别与预先吸附的大鼠和人（P)RR结合后具有酶活性。它们的分子活性分别为4.1 h⁻¹（与人肾素原与人（P)RR的结合（3.8 h⁻¹）几乎相似）和0.98 h⁻¹（比大鼠肾素原与r(P)RR的结合低约2倍）。因此，这些结果表明肾素与（P)RR结合具有物种特异性，这可能有助于阐明肾素与受体结合的生化特性。
