Sabinski F, Wosiewitz U, Leuschner U
Institute of Medical Physics and Biophysics, Westfälische Wilhelms-Universität Münster, Germany.
Eur J Clin Chem Clin Biochem. 1992 Nov;30(11):753-9. doi: 10.1515/cclm.1992.30.11.753.
Native mucin-like complexes were obtained from both pig gallbladder bile and pig gallbladder wall mucus by precipitation, centrifugation, and gel permeation chromatography. Crude preparations by either dialysis (native mucus from bile, and native gallbladder wall mucus) or by precipitation (crude total bile mucus, and lipomucoid) were purified by gel permeation chromatography on Sephacryl S-300HR and Sephacryl S-500HR (Pharmacia). The elution profiles obtained with a reversibly denaturing and detergent-containing eluent showed the same pattern for all samples, although the amounts of the four main fractions differed somewhat. The excluded fraction with the highest carbohydrate portion had an apparent M(r) > 10(7). This fraction and the following included lipomucoid (in physiological solution tightly bound to fraction I), and an eluent-insoluble mucus portion from all samples were characterized by determination of the protein concentration, carbohydrates, sialic acids, and lipids, using standard methods. Sugar analysis was performed by gas-liquid chromatography. Human gallbladder bile was subjected to the same procedures of mucus precipitation and separation. Human gallbladder bile mucus showed identical behaviour to that of pig gallbladder bile mucus, and showed a very similar elution pattern in gel chromatography.
通过沉淀、离心和凝胶渗透色谱法从猪胆囊胆汁和猪胆囊壁黏液中获得天然黏蛋白样复合物。通过透析(胆汁中的天然黏液和天然胆囊壁黏液)或沉淀(粗制总胆汁黏液和脂类黏蛋白)得到的粗制品,在Sephacryl S - 300HR和Sephacryl S - 500HR(Pharmacia)上进行凝胶渗透色谱法纯化。使用含可逆变性剂和去污剂的洗脱液获得的洗脱曲线显示,所有样品的模式相同,尽管四个主要组分的量略有不同。碳水化合物含量最高的排阻组分的表观相对分子质量(M(r))> 10⁷。使用标准方法通过测定蛋白质浓度、碳水化合物、唾液酸和脂质,对该组分以及随后包含的脂类黏蛋白(在生理溶液中与组分I紧密结合)和所有样品中不溶于洗脱液的黏液部分进行了表征。糖分析通过气液色谱法进行。人胆囊胆汁经过相同的黏液沉淀和分离程序。人胆囊胆汁黏液与猪胆囊胆汁黏液表现出相同的行为,并且在凝胶色谱中显示出非常相似的洗脱模式。