CAJ | 학술논문

　　目的验证蔗糖密度梯度超速离心法提取脑组织脂筏的有效性。方法用蔗糖密度梯度超速离心法提取小鼠脑组织脂筏，采用免疫印迹法、双酶胆固醇检测法结合光散射度分析样品的脂筏属性，采用液质联用蛋白组学技术和生物信息学手段，对脂筏中的蛋白质细胞定位进行分析。结果超速离心法提取的脑组织脂筏具有典型的高散色度、高胆固醇和高表达Flotillin-1的脂筏特性；液质联用蛋白组学分析从脂筏样本中鉴定出647种蛋白质，这些蛋白质细胞定位大多是细胞膜、内质网、细胞骨架和细胞浆等常见的脂筏蛋白来源，这种脂筏样品是含有杂质的混合物，鉴定出的647种总蛋白中，有21%是细胞核、线粒体和核糖体等非脂筏来源蛋白。结论超速离心法是提取脑组织脂筏的有效方法，但应用中要考虑杂蛋白的影响。
　　목적험증자당밀도제도초속리심법제취뇌조직지벌적유효성。방법용자당밀도제도초속리심법제취소서뇌조직지벌，채용면역인적법、쌍매담고순검측법결합광산사도분석양품적지벌속성，채용액질련용단백조학기술화생물신식학수단，대지벌중적단백질세포정위진행분석。결과초속리심법제취적뇌조직지벌구유전형적고산색도、고담고순화고표체Flotillin-1적지벌특성；액질련용단백조학분석종지벌양본중감정출647충단백질，저사단백질세포정위대다시세포막、내질망、세포골가화세포장등상견적지벌단백래원，저충지벌양품시함유잡질적혼합물，감정출적647충총단백중，유21%시세포핵、선립체화핵당체등비지벌래원단백。결론초속리심법시제취뇌조직지벌적유효방법，단응용중요고필잡단백적영향。
Objective To verify the validity of the sucrose density gradient ultracentrifugation method for lipid rafts from cerebral cortex. Methods Extract lipid rafts from cerebral cortex in mouse were extracted by the sucrose density gradi-ent ultracentrifugation method. The properties of lipid rafts were detected by Western blotting method, double enzyme and light scattering methods. HPLC MS/MS proteomics and bioinformatics were used to locate proteins of lipid rafts in cells. Re-sults Lipid rafts from cerebral cortex were provided with the model properties of lipid rafts such as high light scattering and cholesterol and high expression of Flotillin-1. HPLC MS/MS proteomics identified total 647 proteins. Most of these pro-teins were from plasma membrane, endoplasmic reticulum, cytoskeleton and cytosol, however, there were 21% proteins among total 647 proteins were from nucleus, mitochondria and ribosomes. Conclusion The sucrose density gradient ultra-centrifugation method is a effective method to extract lipid rafts from cerebral cortex, however, the properties of mixture should be considered.