KANE R E
J Cell Biol. 1965 Apr;25(1):SUPPL:137-44. doi: 10.1083/jcb.25.1.137.
Previous investigations have shown that the mitotic apparatus (MA) can be isolated from dividing sea urchin eggs in water buffered at pH 5.6 and that the addition of 1 M hexanediol to the solution raises the usable pH to 6.4. Long chain glycols appeared to be much more effective than related compounds in increasing the stability of the MA, and the aim of the investigations reported here was to determine the basis of this specificity. These experiments show that this impression of specificity is misleading and that under suitable experimental conditions a variety of compounds can be substituted for the glycols. A number of alcohols will duplicate the action of the glycols in stabilizing the MA at pH 6.4, but they must be used at a similar per cent concentration rather than at a similar molar concentration. Increases in the concentration of alcohol or glycol allow isolation at more alkaline pH values, and a pH-concentration relation for the stability of the MA, covering the range from pH 5.6-8, has been determined. These results indicate that the action of these compounds in stabilizing the mitotic apparatus is non-specific and is similar to their effects on the solubility of proteins. The isolation and stabilization of the mitotic apparatus can thus be viewed as a function of the solubility properties of its constituent proteins, opening a variety of new experimental approaches to this problem.
先前的研究表明,有丝分裂器(MA)可以从处于pH 5.6缓冲水中正在分裂的海胆卵中分离出来,并且向溶液中添加1 M己二醇可将可用pH提高到6.4。在提高有丝分裂器的稳定性方面,长链二醇似乎比相关化合物有效得多,本文报道的研究目的是确定这种特异性的基础。这些实验表明,这种特异性的印象具有误导性,并且在合适的实验条件下,多种化合物可以替代二醇。许多醇在pH 6.4时能在稳定有丝分裂器方面发挥与二醇相同的作用,但必须以相似的百分比浓度而非相似的摩尔浓度使用。醇或二醇浓度的增加使得在更碱性的pH值下也能进行分离,并且已经确定了有丝分裂器稳定性的pH-浓度关系,其范围涵盖pH 5.6至8。这些结果表明,这些化合物在稳定有丝分裂器方面的作用是非特异性的,并且与其对蛋白质溶解度的影响相似。因此,有丝分裂器的分离和稳定可被视为其组成蛋白质溶解性的一种功能,为解决这个问题开辟了各种新的实验方法。