Detection and purification of materials affecting cell function from the classified barley flour using a microtubule assembly system

使用微管组装系统检测和纯化分级大麦粉中影响细胞功能的物质

• 批准号: 06680048
• 负责人: DOI Hiroshi
• 金额: $ 1.41万
• 依托单位: Mukogawa Women's University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06680048/
• 关键词: Microtubule; Tubulin; Barley; Barley Flour; Classification; Cell Funciton; 微小管形成; GTPase活性

Barley is one of the major crop produced in the world. The majority of barley produced is used as animal feed. The use of barley is therefore very different from other grain crops. The reason for this is that the grain is much harder than that of rice or wheat. Consequently, the useage of crop is limited with ocnsequent decrease in economic value. Barley is usefull as a foodstuff, because it contains high levels of protein, dietary fibers and minerals. Recently a new method for the milling of barley grain by a tangential scraping action. We have been attampting to isolate biologically active materials from classified barley flour.It is well-known that tubulin exists in eukaryotic cell widely, and that it forms microtubules by self assembly. Microtubules take part in various cell functins. When tubulin forms microtubules, GTPase activity is observed. It means that materials affecting GTPase activity of tubulin take part in the cell functions.In this study, the purification of the material affecting GTPase activity of tubulin is described. The materials was obtained from the classifed barley flour produced in Japan. The 95-75 % classification flour was used as the starting material for the purification. After cold aceton treatment of the flour, water extract was chromatographed on a Sephacryl S-100HR column. The final preparation obtained from DEAE-Sephadex A-50 ion exchange chromatography and Bio-Gel P-4 gel chromatography was a glycoprotein with a molecular weight of 14,500 dalton containing much aspartic acid and glycine.

大麦是世界主要粮食作物之一。生产的大部分大麦用作动物饲料。因此，大麦的用途与其他粮食作物大不相同。这是因为谷物比大米或小麦硬得多。因此，作物的利用受到限制，经济价值不断下降。大麦是有用的食品，因为它含有高水平的蛋白质，膳食纤维和矿物质。最近提出了一种通过切向刮擦作用研磨大麦的新方法。微管蛋白广泛存在于真核细胞中，它通过自组装形成微管，因此，我们一直致力于从大麦粉中分离具有生物活性的物质。微管参与各种细胞功能。当微管蛋白形成微管时，观察到GTdR活性。这意味着影响微管蛋白GT3活性的物质参与了细胞的功能，本研究对影响微管蛋白GT3活性的物质进行了纯化。材料来自日本生产的分类大麦粉。将95- 75%分级粉用作纯化的起始材料。面粉经冷丙酮处理后，水提取物在Sephacryl S-100 HR柱上层析。经DEAE-Sephadex A-50离子交换层析和Bio-Gel P-4凝胶层析得到的最终产物是一种分子量为14，500道尔顿的糖蛋白，含有大量的天冬氨酸和甘氨酸。