Takahashi S, Miura R, Miyake Y
J Biochem. 1985 Feb;97(2):671-7. doi: 10.1093/oxfordjournals.jbchem.a135103.
We purified, from human kidney, a protein that reacts with rabbit anti-porcine kidney renin binding protein (RnBP) antiserum by trapping with porcine kidney renin. The purified preparation showed a single protein peak on gel filtration by high performance liquid chromatography (HPLC) and two protein bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The latter two kinds of protein were identified as the porcine renin and human kidney protein from their electrophoretic mobilities and reactivity toward rabbit anti-porcine kidney renin and RnBP antisera. The molecular weights of the purified preparation and the human kidney protein were estimated to be 56,000 by HPLC and 43,000 by SDS-PAGE, respectively. The specific activity of porcine renin in the purified preparation was 8.6 mg angiotensin I per mg of protein per h at 37 degrees C and pH 6.5. This specific activity was about one-fifth that of free porcine renin. Therefore, it is suggested from the reactivity toward the anti-porcine RnBP antiserum and inhibitory action toward porcine renin that the human kidney protein is RnBP and that the human RnBP is purified as a complex with porcine renin.
我们通过用猪肾肾素进行捕获,从人肾中纯化出一种能与兔抗猪肾肾素结合蛋白(RnBP)抗血清发生反应的蛋白质。在高效液相色谱(HPLC)的凝胶过滤中,纯化制剂显示出单一的蛋白峰,而在十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)中则显示出两条蛋白带。根据这两种蛋白的电泳迁移率以及它们对兔抗猪肾肾素和RnBP抗血清的反应性,鉴定出后两种蛋白分别为猪肾素和人肾蛋白。通过HPLC和SDS-PAGE分别估计,纯化制剂和人肾蛋白的分子量分别为56,000和43,000。在37℃和pH 6.5条件下,纯化制剂中猪肾素的比活性为每毫克蛋白每小时8.6毫克血管紧张素I。该比活性约为游离猪肾素的五分之一。因此,从其对抗猪RnBP抗血清的反应性以及对猪肾素的抑制作用来看,提示人肾蛋白为RnBP,并且人RnBP是作为与猪肾素的复合物被纯化出来的。