Carlsberg Laboratory, Copenhagen, Denmark.
J Gen Physiol. 1927 Mar 7;8(6):543-99. doi: 10.1085/jgp.8.6.543.
Introduction. A description is given of the principle followed in the experimental determination of the ionisation of egg albumin, its capacity to combine with acids and bases. Egg albumin is regarded as an ampholyte, and in accordance with J. N. Brønsted's definition of acids and bases, ampholytes are considered as substances capable of both taking up and giving off hydrogen ions. The theoretical treatment of the capacity of ampholytes to combine with acids (and bases) has been carried out on this basis. Section A. Several experimental series are noted, comprising the determination of the activity coefficient of the hydrogen ion (fH) in ammonium chloride solutions of different concentration. Section B. The general method of experimental determination of the ionisation (capacity to combine with adds and bases) of egg albumin in ammonium chloride and potassium chloride solutions is briefly described, and the results of the experiments are compared. Section C. 1). In a brief theoretical survey we have suggested that distinction should be made between isoelectric and isoionic reaction of an ampholyte, the former defined as the hydrogen ion activity (value of paH) at which the mean valency of the ampholyte is 0, the latter as the hydrogen ion activity at which the quantity of acid or base combined with the ampholyte is 0; or, as we prefer to express it, the hydrogen ion activity at which the specific hydrogen ionisation of the ampholyte is 0. If the ampholyte does not combine with other ions than the hydrogen ion, then isoelectric and isoionic reaction coincide. Isoionic reaction is determined by acid-combining experiments. The principle of this determination is briefly described. A theoretical investigation of the alteration with salt concentration of both isoelectric (isoionic) reaction and the shape and direction of the ionisation curves is made, with regard to ampholytes capable only of combining with hydrogen ions, on the basis of the Debye-Hückel formulae and Linderstrøm-Lang's theory for the ionisation of polyvalent ampholytes of simple type. It is shown that the salt effect, in accordance with the theory, and in qualitative agreement with the experiments, consists in a turning of the ionisation curves, indicating the relation between the quantity of combined acid (specific hydrogen ionisation) and paH, and the turning of the curves, which leaves the isoelectric reaction unaltered, tends in such a direction that the quantity of combined acid at constant ampholyte concentration and constant pan increases with increasing salt concentration. The possibility of chemical combining of other ions than the hydrogen ion is discussed. 2). Following on 1), a brief survey of the experimental results is given. 3). The isoionic reaction is found from the experimental material and proved to be independent of the ammonium chloride concentration. As the mean of all determinations we have pa(H) (0) = 4.898 (isoionic reaction). The difference between this value and that formerly found for ammonium sulphate solutions (4.844) is discussed. 4). Finally, on the basis of the theory in Section 1), some simple calculations of the ionisation curves for egg albumin are made, and it appears that the theory can reproduce the experimental results in a rough quantitative way when we assume that the egg albumin has a radius of 2.21.10(-7) cm. (answering to a molecular weight of 35,000 in aqueous solution), and contains 30 acid and base groups.
简介。本文介绍了实验测定卵白蛋白电离度、与酸碱结合能力的原理。卵白蛋白被视为两性电解质,根据 J. N. 布仑斯特对酸碱的定义,两性电解质被认为是既能接受又能释放氢离子的物质。在此基础上,对两性电解质与酸碱结合的能力进行了理论处理。A 节。本文记录了几个实验系列,包括测定不同浓度氯化铵溶液中氢离子活度系数(fH)。B 节。简要描述了在氯化铵和氯化钾溶液中实验测定卵白蛋白电离度(与酸碱结合能力)的一般方法,并比较了实验结果。C 节。1)。在简要的理论综述中,我们建议区分两性电解质的等电和等离子反应,前者定义为两性电解质平均价数为 0 时的氢离子活度(paH 值),后者定义为与两性电解质结合的酸或碱量为 0 时的氢离子活度;或者,正如我们更愿意表达的那样,是两性电解质的特定氢离子化度为 0 时的氢离子活度。如果两性电解质除了氢离子外不与其他离子结合,那么等电和等离子反应是重合的。等离子反应由结合酸的实验来确定。简要描述了这种测定的原理。基于德拜-休克尔公式和林德斯特罗姆-朗对单价简单多价两性电解质电离的理论,对仅能与氢离子结合的两性电解质的盐浓度对等电(等离子)反应以及电离曲线的形状和方向的变化进行了理论研究。结果表明,盐效应与理论相符,与实验定性一致,表现为电离曲线的转向,表明结合酸的量(特定氢离子化度)与 paH 之间的关系,以及不改变等电反应的曲线转向,使得在恒定两性电解质浓度和恒定 pan 下与酸结合的量随盐浓度的增加而增加。讨论了其他离子(除氢离子外)与两性电解质化学结合的可能性。2)。继 1)之后,简要综述了实验结果。3)。从实验材料中得出等离子反应,并证明其与氯化铵浓度无关。作为所有测定的平均值,我们得到 pa(H)(0)=4.898(等离子反应)。与以前在硫酸铵溶液中发现的值(4.844)的差异进行了讨论。4)。最后,根据第 1 节中的理论,对卵白蛋白的电离曲线进行了一些简单的计算,结果表明,当假设卵白蛋白的半径为 2.21.10(-7)cm(在水溶液中对应于 35,000 分子量),并包含 30 个酸碱基团时,该理论可以以粗略的定量方式再现实验结果。