纯化和表征的一种新型抑制剂的尿激酶从人类尿液。定量和初步表征等离子体。

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树桩,Thienpont M, Collen D

纯化和表征的一种新型抑制剂的尿激酶从人类尿液。定量和初步表征等离子体。

生物化学杂志。1986年9月25日,261 (27):12759 - 66。

PubMed ID

3091604 (在PubMed

]

文摘

Urokinase-related人类尿液中蛋白质发生主要是1:1复杂的尿激酶抑制剂(树桩,d . C。Thienpont, M。,Collen d(1986)生物。261年化学,1267 - 1273)。与这个urokinase-urokinase BALB / c小鼠免疫抑制剂复杂和脾脏细胞与小鼠骨髓瘤细胞融合,导致生产单克隆抗体杂交瘤细胞。三个抗体反应与尿激酶是利用复杂的但不是开发一个敏感(0.5 ng / ml)尿激酶抑制剂的酶联免疫吸附试验,由色谱用于监测其净化chelate-Sepharose锌,伴刀豆球蛋白A-Sepharose, SP-Sephadex 50,和交联葡聚糖g - 100。明显的同质糖蛋白先生50000年获得的收益率40微克/升的尿液和净化320倍。一毫克的纯化蛋白抑制35000 IU的30分钟内尿激酶在37度c。这种蛋白质纯化urokinase-urokinase抑制剂复杂和免疫相关抑制剂的分离部分通过亲核的分裂。免疫有别于所有已知的蛋白酶抑制剂,包括组织类型的内皮细胞衍生快速抑制纤溶酶原激活物,尿激酶和蛋白酶nexin的胎盘抑制剂。在电泳的蛋白质与beta-mobility迁移。抑制尿激酶发生二级速率常数(k)的8×10 (3)m - 1 s - 1的缺失和9 X 10 (4) m - 1 s - 1在50 IU的肝素/毫升。 The urokinase inhibitor was inactive towards single-chain urokinase-type plasminogen activator and plasmin, but it inhibited two-chain tissue-type plasminogen activator with a k below 10(3) M-1 s-1 and thrombin with a k of 4 X 10(4) M-1 s-1 in the absence and 2 X 10(5) M-1 s-1 in the presence of heparin. The concentration of this urokinase inhibitor in plasma from normal subjects determined by immunoassay was 2 +/- 0.7 micrograms/ml (mean +/- S.D., n = 25). The protein purified from plasma by immunoabsorption had the same Mr, amino acid composition, and immunoreactivity as the urinary protein. Furthermore, when urokinase was added to plasma, time-dependent urokinase-urokinase inhibitor complex formation was observed at a rate similar to that observed for the inhibition of urokinase by the purified inhibitor from urine. This urokinase inhibitor, purified from human urine, most probably represents a new plasma protease inhibitor.

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药物靶点

药物	目标	类	生物	药理作用	行动
Tenecteplase	纤溶酶原激活物抑制剂2	蛋白质	人类	未知的	不可用	细节