Jabbal I, Kells D I, Forstner G, Forstner J
Can J Biochem. 1976 Aug;54(8):707-16. doi: 10.1139/o76-102.
Goblet cell mucin (GCM) has been purified for the first time from mucosal scrapings of human small intestine. Proteolytic enzymes and organic solvents were avoided during the isolation procedure. The mucin was purified by Sepharose 4B and 2B column chromatography of high-speed supernatant fractions. The most purified fraction was compared with rat intestinal GCM. The two were similar with respect to chemical composition, antigenic features, and polyacrylamide disc gel electrophoresis. The major chemical differences included a higher hexosamine-fucose and hexosamine-sialic acid ratio in human mucin. The two mucins showed strong concentration dependence in sedimentation velocity studies. Human mucin at a concentration of 0.2 to 1.5 mg protein per millilitre gave multiple associated peaks with variable So values (10.8-36.6). Rat mucin, in contrast, gave a constant (although polydisperse) pattern with So = 15.15. To explore these differences both mucins were stained with periodic acid - Schiff reagent and subjected to band ultracentrifugation at concentrations of 0.6-1.9 mug protein per millilitre. At this low concentration, rat mucin did not change in its sedimentation characteristics. In contrast, human GCM produced a single peak with So = 37.9. Thus dilution abolished polydispersity in the human but not the rat mucin, suggesting that intermolecular bonding forces in the human mucin are weaker.
杯状细胞粘蛋白（GCM）首次从人小肠黏膜刮屑中纯化得到。在分离过程中避免使用蛋白水解酶和有机溶剂。通过对高速上清液组分进行琼脂糖4B和2B柱色谱法来纯化粘蛋白。将纯化程度最高的组分与大鼠肠GCM进行比较。两者在化学组成、抗原特性和聚丙烯酰胺圆盘凝胶电泳方面相似。主要的化学差异包括人粘蛋白中己糖胺 - 岩藻糖和己糖胺 - 唾液酸的比例更高。在沉降速度研究中，两种粘蛋白均表现出强烈的浓度依赖性。浓度为每毫升0.2至1.5毫克蛋白质的人粘蛋白产生多个具有可变S₀值（10.8 - 36.6）的相关峰。相比之下，大鼠粘蛋白呈现出恒定（尽管多分散）的模式，S₀ = 15.15。为了探究这些差异，两种粘蛋白均用高碘酸 - 席夫试剂染色，并在每毫升0.6 - 1.9微克蛋白质的浓度下进行区带超速离心。在这种低浓度下，大鼠粘蛋白的沉降特性没有变化。相比之下，人GCM产生了一个S₀ = 37.9的单峰。因此，稀释消除了人粘蛋白的多分散性，但大鼠粘蛋白没有，这表明人粘蛋白中的分子间结合力较弱。
