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Studies on the structure of carbohydrate moiety in ovoimucin.

卵粘蛋白碳水化合物部分结构的研究。

基本信息

批准号：
61560321
负责人：
ITOH Takatoshi
金额：
$ 1.28万
依托单位：
TOHOKU UNIVESITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61560321/
关键词：
Ovomucin Egg white Electrophoresis Oligosaccharide

项目摘要

Ovomucin is a glycoprotein fraction of egg white characterized by high molecular weight and a contributor to the viscous property of egg white. A study was made on the electrophoretic analysis, fractionation and characterization of chalaza, and ovomucin from thick and thin albumen of the hen's egg. The chalaza and ovomucin were confirmed to be composed of three components which the molecular weights were estimated to be 150,000, 220,000 and 400,000 daltons, and named <alpha>1, <alpha>2 and <beta> respectively. They were fractionated into carbohydrate poor (<alpha>1 and <alpha>2 mixture) and rich (<beta>) components by gel filtration in the presence of SDS and 2-mercaptoethanol. The <beta>-component contains much higher concentrations of carbohydrates than <alpha>-component, except for mannose. The total content of carbohydrate amount to about 50 %, whereas that of <alpha>-component is about 11 %. The carbohydrate chain in <alpha>-ovomucin was considered asparagine-binding N-glycosidic type, whereas that in the <beta>-ovomucin is O-grycosidic type linked to serine or threonine.The <beta>-ovomucin was collected and hydrolyzed by proteinases. The carbohydrate chains linked to the glycopeptide were released by alkaline borohydrate treatment. Three acidic oligosaccharides were obtained by fractionation with DEAE-Sephadex ion exchange chromatography. Carbohydrate compositions were analyzed by gas chromatography, and followed by sequence analysis with direct MS of the permethylated oligosaccharides, GC-MS analysis of pertially methylated derivatives and by anomer analysis with ^<13>C-NMR. The chemical structures identified were Gal-<beta>-1-3-GalNAc-6-SO_3H, NANA-<alpha>-2-3Gal-<beta>-1-3-GalNAc-6-SO_3H, and NANA-<alpha>-2-3-Gal-<beta>-1-3-[NANA-<alpha>-2-3-]-GalNAc.

卵粘蛋白是蛋白白色的一种糖蛋白组分，其特征在于高分子量，并且是蛋白白色粘性的贡献者。对鸡蛋粗蛋白和稀蛋白中的合点蛋白和卵粘蛋白进行了电泳分析、分离和性质研究。结合点和卵粘蛋白由3种组分组成，分子量分别为150，000、220，000和400，000道尔顿，分别命名为<alpha>1、<alpha>2和<beta>。在SDS和2-巯基乙醇存在下，通过凝胶过滤将它们分级为贫碳水化合物（<alpha>1和<alpha>2混合物）和富碳水化合物（<beta>）组分。除甘露糖外，α<beta>-组分所含的碳水化合物浓度远高于<alpha>α-组分。碳水化合物的总含量约为50%，而非碳水化合物的总含量<alpha>约为11%。α<alpha>-卵粘蛋白的糖链是天冬酰胺结合的N-糖苷型，而α<beta>-卵粘蛋白的糖链是O-糖苷型，与丝氨酸或苏氨酸连接<beta>。通过碱性硼水合物处理释放与糖肽连接的碳水化合物链。经DEAE-Sephadex离子交换层析分离得到三种酸性寡糖。通过气相色谱法分析碳水化合物组成，然后通过全甲基化寡糖的直接MS进行序列分析，部分甲基化衍生物的GC-MS分析和通过1HC-NMR进行端基异构体分析<13>。化学结构鉴定为Gal-<beta>1 - 3-GalNAc-6-SO_3H、NANA<alpha>-2 - 3Gal-<beta>1 - 3-GalNAc-6-SO_3H和NANA<alpha>-2 - 3-Gal-<beta>1 - 3-[NANA-<alpha>2 - 3-]-GalNAc。