MEASUREMENT OF URINARY KALLIKREIN ACTIVITY - SPECIES-DIFFERENCES IN KININ PRODUCTION
MEASUREMENT OF URINARY KALLIKREIN ACTIVITY - SPECIES-DIFFERENCES IN KININ PRODUCTION
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DOI：
10.1016/0304-4165(81)90262-2
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
MENARD, J
中科院分区：
文献类型：
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作者：
ALHENCGELAS, F;MARCHETTI, J;MENARD, J
建立了一种灵敏、特异的激肽放射免疫分析方法，可检测1.5 pg缓激肽或3 pg赖氨酰缓激肽。在0.6-ml孵育液中用10 pg缓激肽或15 pg赖氨酸-缓激肽获得标准曲线中50%的位移。抗缓激肽的抗血清能很好地识别赖氨酸-缓激肽和甲硫氨酰-赖氨酰-缓激肽（Met-lys-缓激肽），但与缓激肽片段的交叉反应率为0.4%或更低。激肽原交叉反应仅0.2%。用放射免疫法和几种激肽原测定人和大鼠尿激肽释放酶活性。通过放射免疫测定法在2种不同的系统（聚丙烯酰胺凝胶电泳和羧甲基纤维素色谱）中对大鼠或人尿液水解底物产生的肽进行了表征。两种尿液产生的激肽不同：人类尿液产生的激肽像赖氨酸缓激肽一样迁移，而大鼠尿液产生的激肽像缓激肽一样迁移。这证明了大鼠和人尿液中激肽形成酶之间特异性的差异。
A sensitive, specific radioimmunoassay for kinins was developed, which is able to detect 1.5 pg bradykinin or 3 pg lysyl-bradykinin (Lys-bradykinin). Fifty percent displacement in the standard curve was obtained with 10 pg bradykinin or 15 pg Lys-bradykinin in 0.6-ml incubates. The antisera, raised against bradykinin, recognized well Lys-bradykinin and methionyl-lysyl-bradykinin (Met-lys-Bradykinin), but cross-reacted 0.4% or less with bradykinin fragments. Kininogen cross-reacted only 0.2%. The radioimmunoassay and kininogen from several species were used in the measurement of human and rat urinary kallikrein activity. The peptide generated by hydrolysis of the substrates by rat or human urines was characterized by radioimmunoassay in 2 different systems: polyacrylamide gel electrophoresis and carboxymethyl cellulose chromatography. Both urines did not produce the same kinin: the kinin produced by human urine migrated like Lys-bradykinin, whereas the kinin produced by rat urine migrated like bradykinin. This gives evidence of differences in the specificity between kinin-forming enzymes in rat and human urines.