Isaacs W B, Shaper J H
Endocrinology. 1985 Oct;117(4):1512-20. doi: 10.1210/endo-117-4-1512.
We have recently reported on the isolation and characterization of the predominant protein of canine prostatic fluid and the subsequent identification of this protein as a proteolytic enzyme. In this report, we described the preparation and characterization of a rabbit antiserum against this secretory protease which was purified from canine prostatic fluid by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Specificity of the antiserum has been established by immunoblot analysis and immunoprecipitation of the secretory protease present in canine prostatic fluid and tissue homogenates. The antiserum was used to localize (by immunofluorescence) and quantitate (by single radial immunodiffusion) the secretory protease in prostatic tissue obtained from intact and castrated dogs. These analyses can be summarized as follows: the secretory protease is strictly localized in the apical portions of the columnar epithelial cells in prostates of intact dogs, where it comprises almost 60% of the total Triton X-100-soluble protein; and the level of secretory protease decreases at least 500-fold in prostates of castrated dogs, demonstrating the androgen-dependent nature of this enzyme. Proteins immunologically related to the secretory protease were detected at low levels in the pancreas and salivary glands of the dog. Such proteins were not detectable in canine liver, kidney, vas deferens, testis, epididymis, lung, spleen, skin, small intestine, or sartorius muscle, suggesting a high degree of prostatic specificity for the secretory protease. These results demonstrate that the canine prostate is highly specialized with respect to its androgen-dependent synthesis and secretion of this secretory protease and that the antiserum specific for this protein can be used to assess canine prostatic function under varying hormonal conditions.
我们最近报道了犬前列腺液中主要蛋白质的分离与特性鉴定，随后将该蛋白质鉴定为一种蛋白水解酶。在本报告中，我们描述了一种兔抗血清的制备与特性鉴定，该抗血清针对的是通过制备性十二烷基硫酸钠-聚丙烯酰胺凝胶电泳从犬前列腺液中纯化得到的这种分泌性蛋白酶。通过对犬前列腺液和组织匀浆中存在的分泌性蛋白酶进行免疫印迹分析和免疫沉淀，确定了抗血清的特异性。该抗血清用于（通过免疫荧光）定位和（通过单向放射免疫扩散）定量完整和去势犬前列腺组织中的分泌性蛋白酶。这些分析结果可总结如下：分泌性蛋白酶严格定位于完整犬前列腺柱状上皮细胞的顶端部分，在那里它占总 Triton X - 100 可溶性蛋白的近 60%；去势犬前列腺中分泌性蛋白酶的水平至少降低 500 倍，表明该酶具有雄激素依赖性。在犬的胰腺和唾液腺中检测到与分泌性蛋白酶免疫相关的蛋白质，但含量较低。在犬的肝脏、肾脏、输精管、睾丸、附睾、肺、脾脏、皮肤、小肠或缝匠肌中未检测到此类蛋白质，这表明分泌性蛋白酶具有高度的前列腺特异性。这些结果表明，犬前列腺在雄激素依赖性合成和分泌这种分泌性蛋白酶方面高度特化，并且针对该蛋白质的特异性抗血清可用于评估不同激素条件下的犬前列腺功能。
