Leach B S, Collawn J F, Fish W W
Biochemistry. 1980 Dec 9;19(25):5734-41. doi: 10.1021/bi00566a011.
The influence of the presence of oligosaccharide branches was examined with respect to the behavior of glycopolypeptides in empirical molecular weight estimation methods in the presence of sodium dodecyl sulfate (NaDodSO4). This examination was conducted by comparing the gel chromatographic and gel electrophoretic behaviors in the presence of NaDodSO4 of 13 glycopolypeptides of known chemical and physical properties to those of regular polypeptides. Errors in the gel chromatographic molecular weight for glycopolypeptides in NaDodSO4 varied from -22% to +10% and indicated that the hydrodynamic behavior of the glycopolypeptide--NaDodSO4 complex could not be correlated with the amount of carbohydrate in the glycopolypeptide. NaDodSO4 binding measurements on a number of the glycopolypeptides suggest that the polypeptide moiety binds the nominal weight ratio of NaDodSO4, while the carbohydrate portion exhibits little or no NaDodSO4 binding. As has been reported by others, the polyacrylamide gel electrophoretic behavior of glycopolypeptide--NaDodSO4 complexes yielded abnormally high molecular weight estimates. In general, the error of these estimates diminished with decreasing porosity of the gel; however, each glycopolypeptide behaved in a unique fashion. Treatment of the electrophoretic data by any of several empirical means provided no reliable way to correct for the glycopolypeptides' aberrant behavior.
在十二烷基硫酸钠(NaDodSO4)存在的情况下,针对经验分子量估算方法中糖多肽的行为,研究了寡糖分支存在的影响。通过比较13种已知化学和物理性质的糖多肽与常规多肽在NaDodSO4存在下的凝胶色谱行为和凝胶电泳行为来进行此项研究。糖多肽在NaDodSO4中的凝胶色谱分子量误差在-22%至+10%之间,这表明糖多肽-NaDodSO4复合物的流体动力学行为与糖多肽中碳水化合物的含量无关。对多种糖多肽进行的NaDodSO4结合测量表明,多肽部分结合的NaDodSO4的标称重量比,而碳水化合物部分几乎不结合或不结合NaDodSO4。正如其他人所报道的,糖多肽-NaDodSO4复合物的聚丙烯酰胺凝胶电泳行为给出的分子量估计值异常高。一般来说,这些估计值的误差随着凝胶孔隙率的降低而减小;然而,每种糖多肽的行为都很独特。通过几种经验方法中的任何一种处理电泳数据,都无法提供可靠的方法来校正糖多肽的异常行为。