Nozaki Y, Schechter N M, Reynolds J A, Tanford C
Biochemistry. 1976 Aug 24;15(17):3884-90. doi: 10.1021/bi00662a036.
In the course of a routine investigation of the complex between the erythrocyte membrane protein spectrin and sodium dodecyl sulfate, we observed a large discrepancy between the true Stokes radius (178 A, measured by hydrodynamic methods) and the apparent value derived from gel chromatography (107 A). In attempting to resolve this discrepancy, we have experiments that indicate that all large asymmetric particles may be subject to a similar discrepancy; e.g., native fibrinogne has a true Strokes radius of 108 A, whereas the value derived by column chromatography after calibration with globular proteins is only 71 A. The simplest interpretation is that end-on insertion of asymmetric particles into the gel pores contributes to their retardation. The phenomenon clearly limits the usefulness of gel chromatography as a quantitative measure of the hydrodynamic Stokes radius. Incidental data obtained in the course of this work indicate that spherical viruses may have weak chemical affinity for the porous gel. Chromatography of large proteins in the presence of detergents produced no effects ascribable to absorption of the detergents, but the results suggest a need for further study of possible interaction between detergents and small gel pores.
在对红细胞膜蛋白血影蛋白与十二烷基硫酸钠之间的复合物进行常规研究的过程中，我们观察到真实的斯托克斯半径（通过流体动力学方法测得为178 Å）与凝胶色谱得出的表观值（107 Å）之间存在很大差异。在试图解决这一差异时，我们进行的实验表明，所有大的不对称颗粒可能都存在类似的差异；例如，天然纤维蛋白原的真实斯托克斯半径为108 Å，而在用球状蛋白校准后通过柱色谱得出的值仅为71 Å。最简单的解释是，不对称颗粒以端部插入凝胶孔中导致其滞留。该现象显然限制了凝胶色谱作为流体动力学斯托克斯半径定量测量方法的实用性。在这项工作过程中获得的附带数据表明，球形病毒可能与多孔凝胶具有较弱的化学亲和力。在去污剂存在的情况下对大蛋白进行色谱分析未产生可归因于去污剂吸收的影响，但结果表明需要进一步研究去污剂与小凝胶孔之间可能的相互作用。
