Miller D H, Chao J, Margolius H S
Biochem J. 1984 Feb 15;218(1):37-43. doi: 10.1042/bj2180037.
A method has been developed to measure the relative rate of rat tissue kallikrein synthesis which employs a specific antiserum raised against a purified rat urinary kallikrein. Incorporation of [35S]methionine into kallikrein and protein 20 min after intraperitoneal injection was measured in submaxillary gland, pancreas, kidney and descending colon. Kallikrein content was measured with a direct radioimmunoassay, and kallikrein-specific incorporation of [35S]methionine measured after immunoprecipitation. Kallikrein specific radioactivity (c.p.m./mg of enzyme) was about 100-fold greater than that in total protein in both kidney and colon. In contrast, in pancreas the incorporation into the enzyme was only 5-fold higher than into protein, and in submaxillary gland the incorporation was equivalent. Measured as kallikrein-specific radioactivity relative to total protein radioactivity incorporated in 20 min, kallikrein represents 0.18% of total protein synthesis in the kidney, 0.34% in the pancreas, 0.41% in the colon, but 7.29% in the submaxillary gland. Dietary Na+ restriction increased the relative rate of kallikrein synthesis 1.8-fold in the kidney without a comparable effect in submaxillary gland. In contrast, testosterone increased the relative rate of synthesis 2.3-fold in submaxillary gland, but decreased it in kidney. The data show that endogenous kallikrein synthesis differs markedly in various tissues, and that interventions which are known to change kallikrein content or excretion also change the relative rate of enzyme synthesis.
已开发出一种方法来测量大鼠组织激肽释放酶的相对合成速率,该方法使用针对纯化的大鼠尿激肽释放酶产生的特异性抗血清。腹腔注射20分钟后,在颌下腺、胰腺、肾脏和降结肠中测量[35S]甲硫氨酸掺入激肽释放酶和蛋白质的情况。用直接放射免疫测定法测量激肽释放酶含量,免疫沉淀后测量[35S]甲硫氨酸的激肽释放酶特异性掺入情况。肾脏和结肠中激肽释放酶的比放射性(每分钟计数/毫克酶)比总蛋白中的比放射性大约高100倍。相比之下,在胰腺中,酶的掺入仅比蛋白质高5倍,而在颌下腺中掺入量相当。以相对于20分钟内掺入的总蛋白放射性的激肽释放酶特异性放射性来衡量,激肽释放酶在肾脏中占总蛋白合成的0.18%,在胰腺中占0.34%,在结肠中占0.41%,但在颌下腺中占7.29%。饮食中钠限制使肾脏中激肽释放酶的相对合成速率增加1.8倍,而在颌下腺中没有类似效果。相比之下,睾酮使颌下腺中合成速率增加2.3倍,但使肾脏中的合成速率降低。数据表明,内源性激肽释放酶合成在不同组织中明显不同,并且已知改变激肽释放酶含量或排泄的干预措施也会改变酶的相对合成速率。