Tabak L A, Mirels L, Monte L D, Ridall A L, Levine M J, Loomis R E, Lindauer F, Reddy M S, Baum B J
Arch Biochem Biophys. 1985 Nov 1;242(2):383-92. doi: 10.1016/0003-9861(85)90222-x.
A blood group A+ mucin-glycoprotein was purified from aqueous extracts of rat submandibular glands by sequential chromatography on columns of Sepharose CL-6B and Sephacryl S-300 in urea-containing buffers. Final purification was facilitated by reductive methylation which appeared to release contaminating (hydrophobic) peptides. Homogeneity of the purified mucin was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis at varying concentrations of acrylamide, lectin affinity chromatography, and Western blot analysis. In contrast to previously described preparations, the purified mucin contained only trace amounts of N-acetylglucosamine and aromatic amino acids. In addition, only low levels of basic amino acids were present.
通过在含尿素缓冲液中的Sepharose CL - 6B柱和Sephacryl S - 300柱上进行连续色谱法，从大鼠下颌下腺水提取物中纯化出一种A +血型粘蛋白 - 糖蛋白。还原甲基化有助于最终纯化，它似乎释放出了污染性（疏水）肽。通过在不同丙烯酰胺浓度下的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳、凝集素亲和色谱法和蛋白质印迹分析来确定纯化粘蛋白的同质性。与先前描述的制剂相比，纯化的粘蛋白仅含有痕量的N - 乙酰葡糖胺和芳香族氨基酸。此外，仅存在低水平的碱性氨基酸。
