推荐回答(3个)
After each purification step, the activity of the preparation (in units of enzyme activity) is assayed, the total amount of protein is determined independently, and the ratio of the two gives the specific activity. Activity and total protein generally decrease with each step. Activity decreases because some loss always occurs due to inactivation or nonideal interactions with chromatographic materials or other molecules in the solution.
在每一步提纯之后,分析配置剂(以酶活性为单元)的活性,蛋白质的总量要单独确定,然后由这二者的比例给出确切的活性。活性及蛋白质总量通常会随每一步而减少。活性减少是因为失活,或色析材料/溶液中的其它分子间不理想的相互作用。
Total protein decreases because the objective is to remove as much unwanted or nonspecific protein as possible. In a successful step, the loss of nonspecific protein is much greater than the loss of activity; therefore, specific activity increases even as total activity falls. The data are then assembled in a purification table similar to Table 3–5. A protein is generally considered pure when further purification steps fail to increase specific activity and when only a single protein species can be detected (for example, by electrophoresis).
蛋白质总量的减少是因为要尽可能地移除不想要的或非特定蛋白质的目的。在一个成功的步骤中,非特定的蛋白质的丧失比活性的丧失要大很多;因此,即使总活性下降但特定活性上升。然后在类似表3-5的提纯表中汇集数据。当只检测到单一的蛋白质种类时,该蛋白质通常被认为是纯净的(比如,通过电泳方式)。
For proteins that are not enzymes, other quantification methods are required. Transport proteins can be assayed by their binding to the molecule they transport, and hormones and toxins by the biological effect they produce; for example, growth hormones will stimulate the growth of certain cultured cells. Some structural proteins represent such a large fraction of a tissue mass that they can be readily extracted and purified without a functional assay. The approaches are as varied as the proteins themselves.
因为蛋白质不是酶,需要其他定量方法。转运蛋白可以通过它们所要运送的绑定在它们上的分子来检验,而激素和毒素可以通过它们所产生的生物效应来检验;比如,生长激素将会刺激某些培养细胞的生长。某些结构蛋白表征了一个组织块的大部分,他们很容易萃取并提纯而不需要功能性分析。这些方法随蛋白质本身的不同而各不相同。
净化后每个步骤,活动的
编制(单位酶活性)是检测的
总额蛋白决心独立,
比值的两个给出了具体的活动。活动
和总蛋白降低,一般每
第一步。活动减少一些损失,因为总是发生
由于灭活或非理想的相互作用
色谱材料或其他分子的解决办法。
总蛋白降低,因为目标是
删除了不必要的或特异性蛋白
可能的。在一个成功的一步,损失的非特异性蛋白
远大于损失的活动,因此,
具体活动的增加,尽管总活性下降。那个
数据,然后组装在一个类似的净化表
表3-5 。 A蛋白通常被认为是purewhen进一步净化步骤无法增加的具体
活动时,只有一个物种的蛋白质可
检测(例如,通过电泳) 。
蛋白质不是酶,其他量化
方法是必需的。运输蛋白可
测定它们的结合分子,他们运输,
与激素和毒素的生物效应,他们
生产,例如,生长激素会刺激
增长的某些细胞培养。一些结构性
蛋白质占如此大的几分之组织肿块
他们可以随时提取和纯化无
功能检测。该办法一样有多种theproteins自己。
一楼翻译的不通. 肯定不是自己翻译的. 我想翻可是太长了而且分太少了. 不值得浪费时间. 如果追加分还可以考虑.
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