ISOLATION AND CHARACTERIZATION OF HUMAN CERVICAL-MUCUS GLYCOPROTEINS
ISOLATION AND CHARACTERIZATION OF HUMAN CERVICAL-MUCUS GLYCOPROTEINS
复制标题
人宫颈粘液糖蛋白的分离和表征
DOI：
10.1042/bj2110013
复制
发表时间：
1983-01-01
影响因子：
4.1
通讯作者：
WINGERUP, L
中科院分区：
文献类型：
--
作者：
CARLSTEDT, I;LINDGREN, H;WINGERUP, L
在蛋白酶抑制剂存在下，用6 M-氯化胍从人宫颈妊娠粘液中提取粘液糖蛋白（粘蛋白）。随后通过在CsCl/氯化胍梯度中等密度密度梯度离心实现纯化。纯化的大分子表示为85%的总量和缺乏核酸和蛋白质，判断分析密度梯度离心，圆盘电泳和紫外光谱。沉降速度离心显示离心揭示了一个单一的单峰与s20，w 50.1S在0.2 M-氯化钠和37.0 S在6 M-氯化胍。通过光散射获得的MW为9.7 × 104。106和5.9 ×106分别在0.2 M-NaCl和6 M-氯化胍中。化学分析是典型的上皮粘蛋白。大分子含有. apprx。20%（w/w [wt/wt]）的蛋白质和65%（w/w）的碳水化合物。丝氨酸和苏氨酸构成32 mol/100 mol氨基酸，脯氨酸构成10 mol/100 mol氨基酸。发现的主要糖是N-乙酰葡糖胺（12.8%）、N-乙酰半乳糖胺（9.7%）、半乳糖（18.7%）、唾液酸（15.0%）和岩藻糖（7.5%）。
Mucus glycoproteins (mucins) were extracted from human cervical pregnancy mucus by 6M-guanidinium chloride in the presence of proteinase inhibitors. Purification was subsequently achieved by isopycnic density-gradient centrifugation in CsCl/guanidinium chloride gradients. The purified macromolecules represented .apprx. 85% of the total and were devoid of nucleic acids and proteins, as judged by analytical density-gradient centrifugation, disc electrophoresis and UV spectroscopy. Sedimentation-velocity centrifugation revealed centrifugation revealed a single unimodal peak with s20,w 50.1S in 0.2 M-NaCl and 37.0S in 6M-guanidinium chloride. MW obtained by light-scattering were 9.7 .times. 106 and 5.9 .times. 106 in 0.2 M-NaCl and 6 M-guanidinium chloride, respectively. The chemical analyses were typical of those of epithelial mucins. The macromolecules contained .apprx. 20% (w/w [wt/wt]) of protein, and 65% (w/w) was accounted for as carbohydrate. Serine and threonine constituted 32 mol/100 mol and proline 10 mol/100 mol of the amino acids. The major sugars found were N-acetylglucosamine (12.8%), N-acetylgalactosamine (9.7%), galactose (18.7%), sialic acid (15.0%) and fucose (7.5%).