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猪结肠黏液中高分子量糖蛋白的分离与鉴定

The isolation and characterization of the high-molecular-weight glycoprotein from pig colonic mucus.

作者信息

Marshall T, Allen A

出版信息

Biochem J. 1978 Aug 1;173(2):569-78. doi: 10.1042/bj1730569.

DOI:10.1042/bj1730569
PMID:697737
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1185811/
Abstract
  1. A high-molecular-weight glycoprotein constitutes over 80% by weight of the total glycoprotein from water-soluble pig colonic mucus. 2. It was isolated from from nucleic acid and non-covalently bound protein by nuclease digestion followed by equilibrium centrifugation in a CsCl gradient. 3. The glycoprotein has the following composition by weight: fucose 10.4%; glucosamine 23.9%; galactosamine 8.3%; sialic acid 9.9%; galactose 20.8%; sulphate 3.0%; protein 13.3%; moisture about 10%. 4. The native glycoprotein has the high mol.wt. of 15 X 10(6). 5. Reduction of the native glycoprotein with 2-mercaptoethanol results in a glycoprotein of mol.wt. 6 X 10(6). 6. Pronase digestion removes 29% of the protein (3% of the glycoprotein) but none of the carbohydrate. 7. The molecular weight of the Pronase-digested glycoprotein is 1.5 X 10(6), which is halved to 0.76 X 10(6) on reduction with 2-mercaptoethanol. 8. The contribution of non-covalent interactions, disulphide bridges and the non-glycosylated peptide core to the quaternary structure of the glycoprotein are discussed and compared with the known structure of pig gastric glycoportein.
摘要
  1. 一种高分子量糖蛋白占水溶性猪结肠黏液总糖蛋白重量的80%以上。2. 通过核酸酶消化,随后在CsCl梯度中进行平衡离心,将其与核酸和非共价结合蛋白分离。3. 该糖蛋白的重量组成如下:岩藻糖10.4%;葡糖胺23.9%;半乳糖胺8.3%;唾液酸9.9%;半乳糖20.8%;硫酸盐3.0%;蛋白质13.3%;水分约10%。4. 天然糖蛋白的分子量为15×10⁶。5. 用2-巯基乙醇还原天然糖蛋白会产生分子量为6×10⁶的糖蛋白。6. 链霉蛋白酶消化可去除29%的蛋白质(占糖蛋白的3%),但不会去除任何碳水化合物。7. 链霉蛋白酶消化后的糖蛋白分子量为1.5×10⁶,用2-巯基乙醇还原后减半至0.76×10⁶。8. 讨论了非共价相互作用、二硫键和非糖基化肽核心对糖蛋白四级结构的贡献,并与猪胃糖蛋白的已知结构进行了比较。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/abce/1185811/e15f5549046f/biochemj00482-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/abce/1185811/e15f5549046f/biochemj00482-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/abce/1185811/e15f5549046f/biochemj00482-0219-a.jpg

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