Purification of cholesterol gallstone inhibitory and promoting glycoproteins in bile and its clinical significance

胆汁中胆固醇结石抑制和促进糖蛋白的纯化及其临床意义

• 批准号: 03670383
• 负责人: AOYAMA Nankei
• 金额: $ 1.28万
• 依托单位: Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670383/
• 关键词: Cholesterol gallstone; Cholesterol nucleation; Helix pomatia lectin; Con A lectin; Promoter glycoprotein; Inhibitor glycoprotein; 糖蛋白; Helix pomatia lectia; コレステロ-ル結石形成抑制因子; コレステロ-ル結石形成促進因子; 糖タンパク; Cholesterol Nucleation; Cou A lectin; HPLC

Presence of glycoproteins in human bile which promote or inhibit cholesterol crystal formation in bile has been reported by Holzbach, Groen et al,. Holzbach et al, has reported a new method of cholesterol nucleation and growth assay using spectrophotometer, which can semi-quantify the promoting or inhibitory effects of those glycoproteins on cholesterol crystal formation in bile. We purified glycoproteins by series of procedures using gel chromatography, affinity chromatography(Con A and Helix pomatia lectin columns), and HPLC (ion exchanger column). Promoting and inhibitory activity of purified glycoproteins from human bile was examined by using cholesterol nucleation and growth assay. As the results, glycoproteins which showed promoting activity on cholesterol crystal formation in bile were mainly found in Con A lectin bounding protein fraction and those showed inhibitory activity were found in Helix pomatia lectin bounding fraction.The glycoprotein fraction which exhibited promoting effect on the cholesterol crystal formation in bile contained four different proteins on SDS-PAGE electrophoresis whose molecular weights ranged from 30 kD to 120 Kd. These glycoproteins are under further purification.

霍尔茨巴赫、Groen等报道了人胆汁中存在促进或抑制胆汁中胆固醇晶体形成的糖蛋白。霍尔茨巴赫等报道了一种使用分光光度计的胆固醇成核和生长测定的新方法，该方法可以半定量这些糖蛋白对胆汁中胆固醇晶体形成的促进或抑制作用。我们通过凝胶层析、亲和层析（Con A和Helix pomatia凝集素柱）和HPLC（离子交换柱）等一系列程序纯化糖蛋白。用胆固醇成核生长法测定了人胆汁糖蛋白的促生长和抑制活性。作为结果，对胆汁中胆固醇晶体形成有促进作用的糖蛋白主要存在于Con A凝集素结合蛋白组分中，而对胆固醇晶体形成有抑制作用的糖蛋白主要存在于Hessopomatia凝集素结合蛋白组分中。聚丙烯酰胺凝胶电泳，分子量范围为30 ~ 120 kD。这些糖蛋白正在进一步纯化中。