Evin G, Devin J, Castro B, Menard J, Corvol P
Proc Natl Acad Sci U S A. 1984 Jan;81(1):48-52. doi: 10.1073/pnas.81.1.48.
The complete sequence of the structural gene coding for mouse submaxillary gland renin was recently reported and the amino acid sequence of preprorenin was deduced. This sequence includes a 45-amino acid peptide that corresponds to the prosegment of the renin precursor. To investigate whether peptides related to the renin prosegment are able to inhibit renin activity, we have synthesized four peptides having the following structures: Arg-Ile-Pro-OMe, butyloxycarbonyl(Boc)-Leu-Lys-Lys-Met-Pro-OMe, Boc-Arg-Ile-Pro-Leu-Lys-Lys-Met-Pro-OMe, and Boc-Glu-Arg-Ile-Pro-Leu-Lys-Lys-Met-Pro-OMe (corresponding to amino acids 12-14, 15-19, 12-19, and 11-19, respectively, of the renin prosegment). All four peptides were found to inhibit the activity of pure mouse submaxillary renin on a porcine synthetic tetra-decapeptide in vitro, and the most potent inhibitors exhibited IC50 values in the micromolar range. Enzymatic kinetic studies carried out using peptide 15-19 showed an uncompetitive or a mixed type of inhibition with a Ki value of 2.3 X 10(-6) M at 37 degrees C in 0.5 M citrate/phosphate buffer (pH 6.0).
最近报道了编码小鼠颌下腺肾素的结构基因的完整序列，并推导了前肾素原的氨基酸序列。该序列包括一个与肾素前体的前肽相对应的45个氨基酸的肽段。为了研究与肾素前肽相关的肽是否能够抑制肾素活性，我们合成了具有以下结构的四种肽：精氨酸-异亮氨酸-脯氨酸-甲氧基，叔丁氧羰基（Boc）-亮氨酸-赖氨酸-赖氨酸-甲硫氨酸-脯氨酸-甲氧基，Boc-精氨酸-异亮氨酸-脯氨酸-亮氨酸-赖氨酸-赖氨酸-甲硫氨酸-脯氨酸-甲氧基，以及Boc-谷氨酸-精氨酸-异亮氨酸-脯氨酸-亮氨酸-赖氨酸-赖氨酸-甲硫氨酸-脯氨酸-甲氧基（分别对应于肾素前肽的氨基酸12 - 14、15 - 19、12 - 19和11 - 19）。发现所有这四种肽在体外均能抑制纯小鼠颌下肾素对猪合成十四肽的活性，最有效的抑制剂的IC50值在微摩尔范围内。使用肽15 - 19进行的酶动力学研究表明，在37℃、0.5M柠檬酸/磷酸盐缓冲液（pH 6.0）中，其抑制类型为非竞争性或混合型，Ki值为2.3×10^(-6)M。
