Ofosu F, Forstner J, Forstner G
Biochim Biophys Acta. 1978 Nov 1;543(4):476-83. doi: 10.1016/0304-4165(78)90302-1.
Rat intestinal mucin was labelled biologically by intraperitoneal injection of radioactive amino acids and monosaccharides 3--6 h prior to killing, followed by isolation and purification of the mucin from mucosal scrapings. The labelled product was then introduced into intestinal segments of rats under ether anesthesia for periods up to 3 h, removed by washing and assessed for evidence of degradation. In segments containing the pancreatic ducts the total mucin precipitable by cetyltrimethylammonium bromide fell from 80% to 5% in 3 h. At 3 h, chromatography on Sephadex G-100 and Sepharose 4B revealed multiple products, including very small molecular weight fragments deficient in carbohydrate label. With the introduction of neomycin sulfate into the segments to reduce bacterial growth, only two products were found, one corresponding in size to the original mucin and one somewhat smaller, although still in excess of 200,000 daltons. These products occurred independently of the presence of the pancreatic ducts in the segments, and in chronically pancreatectomized rats. The smaller product could not be produced by incubation with trypsin or elastase. Both products were altered antigenically as compared with the original mucin. Both products also retained the same ratio of carbohydrate and protein label as the original. It is concluded that mucins undergo early degradative changes in the intestine which do not involve deglycosylation but which involve partial loss of antigenicity and a fall in molecular weight. The pancreas is not responsible for these changes.
在处死大鼠前3 - 6小时,通过腹腔注射放射性氨基酸和单糖对大鼠肠粘蛋白进行生物标记,随后从粘膜刮片中分离并纯化粘蛋白。然后在乙醚麻醉下将标记产物引入大鼠肠段,持续3小时,通过冲洗去除,评估其降解证据。在含有胰管的肠段中,十六烷基三甲基溴化铵可沉淀的总粘蛋白在3小时内从80%降至5%。3小时时,在葡聚糖凝胶G - 100和琼脂糖4B上进行色谱分析显示有多种产物,包括碳水化合物标记不足的非常小分子量片段。在肠段中加入硫酸新霉素以减少细菌生长,仅发现两种产物,一种大小与原始粘蛋白相对应,另一种稍小,但仍超过200,000道尔顿。这些产物的出现与肠段中胰管的存在无关,在长期胰腺切除的大鼠中也是如此。较小的产物不能通过与胰蛋白酶或弹性蛋白酶孵育产生。与原始粘蛋白相比,这两种产物的抗原性都发生了改变。这两种产物还保留了与原始粘蛋白相同的碳水化合物和蛋白质标记比例。结论是,粘蛋白在肠道中会发生早期降解变化,这种变化不涉及去糖基化,但涉及抗原性的部分丧失和分子量的下降。这些变化与胰腺无关。