Winzor D J, Carrington L E, Harding S E
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland 4072, Australia.
Anal Biochem. 2001 Dec 15;299(2):235-40. doi: 10.1006/abio.2001.5408.
A method is described whereby sedimentation velocity is combined with equilibrium dialysis to determine the net charge (valence) of a protein by using chromate as an indicator ion for assessing the extent of the Donnan redistribution of small ions. The procedure has been used in experiments on bovine serum albumin under slightly alkaline conditions (pH 8.0, I 0.05) to illustrate its application to a system in which the indicator ion and protein both bear net negative charge and on lysozyme under slightly acidic conditions (pH 5.0, I 0.10) to illustrate the situation where chromate is a counterion.
本文描述了一种方法,即通过沉降速度与平衡透析相结合,以铬酸盐作为指示离子来评估小离子的唐南再分布程度,从而确定蛋白质的净电荷(价态)。该方法已用于在微碱性条件(pH 8.0,离子强度0.05)下对牛血清白蛋白的实验,以说明其在指示离子和蛋白质均带净负电荷的体系中的应用,以及在微酸性条件(pH 5.0,离子强度0.10)下对溶菌酶的实验,以说明铬酸盐作为抗衡离子的情况。