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镉离子对凝血酶原吸附到氢氧化铝上的影响,以此作为纯化凝血酶原的一种方法。

The influence of cadmium ions on the adsorption of prothrombin onto A1(OH)3 as a means to purify prothrombin.

作者信息

Devilee P P, Hemker H C, Bas B M

出版信息

Biochim Biophys Acta. 1975 Jan 30;379(1):172-9. doi: 10.1016/0005-2795(75)90019-7.

Abstract
  1. In the presence of CdSO4(1mM),Al(OH)3(1.3% w/v) completely adsorbs the coagulation factors VII, IX, and X from normal plasma, but factor II (prothrombin) is adsorbed for about 50% only.2. A purification procedure for factor II is developed, using Al(OH)3 adsorption in the presence of Cd2+ as a first step and using column chromatography only once. A 750-fold purification is obtained at a 24% yield. 3. Comparison of the prothrombin thus obtained, with prothrombin isolated by the method of Kisiel and Hanahan (Biochim. Biophys. Acta (1973) 304, 103-113) does not show significant differences in amino acid composition, N-terminal amino acid, molecular weight or immunological properties. 4. Comparison of the two prothrombin preparations in a thrombin-generating system shows that although the final yield of thrombin from a given amount of prothrombin in both preparations is the same, the initial velocity of thrombin formation from our preparation is comparable to that of native prothrombin, whereas the other preparation is converted significantly slower.
摘要
  1. 在硫酸镉(1毫摩尔)存在的情况下,氢氧化铝(1.3%重量/体积)能完全吸附正常血浆中的凝血因子VII、IX和X,但凝血因子II(凝血酶原)仅被吸附约50%。2. 开发了一种凝血因子II的纯化程序,第一步是在镉离子存在的情况下用氢氧化铝吸附,且仅使用一次柱色谱法。纯化倍数达750倍,产率为24%。3. 将如此获得的凝血酶原与通过基西尔和哈纳汉方法(《生物化学与生物物理学报》(1973年)304卷,103 - 113页)分离得到的凝血酶原进行比较,结果表明在氨基酸组成、N端氨基酸、分子量或免疫学特性方面没有显著差异。4. 在凝血酶生成系统中对两种凝血酶原制剂进行比较表明,虽然两种制剂中给定数量的凝血酶原最终产生的凝血酶产量相同,但我们制备的制剂中凝血酶形成的初始速度与天然凝血酶原相当,而另一种制剂的转化速度明显较慢。

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