Isolation and characterization of rat plasma glandular kallikrein.
Isolation and characterization of rat plasma glandular kallikrein.
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大鼠血浆腺激肽释放酶的分离和表征。
DOI：
10.1016/0006-2952(85)90099-1
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发表时间：
1985
影响因子：
5.8
通讯作者：
I. Pinto
中科院分区：
文献类型：
--
作者：
J. Masferrer;R. Albertini;H. Croxatto;P. García;I. Pinto
已开发出一种通过使用琼脂糖凝胶-抑肽酶亲和层析和用对氨基苯甲脒洗脱酶来纯化大鼠血浆中存在的腺激肽释放酶的方法。分离的酶从低分子量激肽原 II 中释放激肽（sp. act. 14 ng 激肽/min × mg），并从底物 S-2266 中释放对硝基苯胺 (pNA)（sp. act. 1.23 nmoles pNA/min × mg）；它被抑肽酶、苯甲脒和大鼠尿抗激肽释放酶抗体抑制，但不被卵类粘蛋白抑制。在聚丙烯酰胺凝胶电泳中，制剂的酶活性与分子量等于尿激肽释放酶（35,000 道尔顿）的两条轻蛋白带相关。使用这种方法，添加到血浆中的[125I]激肽释放酶的回收率为82-88%。正常大鼠血浆中酶的浓度相当于 6.1 ±2.1 (S.D.) ng 激肽释放酶/ml。肾切除大鼠的平均值为 20.0 ± 6.3 (S.D.) ng 激肽释放酶/ml。该增量非常显着（P < 0.001）。我们的结果证实血浆中存在腺激肽释放酶，这是通过其他方法检测到的；他们还证明从血浆中纯化的材料具有酶活性，这表明激肽释放酶可能在控制血液循环中发挥生物学作用。
A method has been developed to purify glandular kallikrein present in rat plasma by using Sepharose-Aprotinin affinity chromatography and elution of the enzyme withp-aminobenzamidine. The isolated enzyme liberated kinins from kininogen II of low molecular weight (sp. act. 14 ng kinins/min × mg) andp-nitroaniline (pNA) from the substrate S-2266 (sp. act. 1.23 nmoles pNA/min × mg); it was inhibited by aprotinin, benzamidine and rat urinary antikallikrein antibody but not by ovomucoid. In polyacrylamide gel electrophoresis, the enzymatic activities of the preparation were associated with two light protein bands of molecular weights equal to that of urinary kallikrein (35,000 daltons). Using this method, the recovery of [125I]kallikrein added to the plasma was 82–88%. The concentration of the enzyme in normal rat plasma was equivalent to 6.1 ±2.1 (S.D.) ng kallikrein/ml. The mean value found in nephrectomized rats was 20.0 ± 6.3 (S.D.) ng kallikrein/ml. This increment was highly significant (P < 0.001). Our results confirm the presence of glandular kallikrein in plasma which had been detected by other methods; they also demonstrate that the material purified from plasma is enzymatically active, suggesting that kallikrein may play a biological role in the control of blood circulation.