重组鸡半胱氨酸蛋白酶抑制剂类淀粉样沉淀分析  

Amyloid precipitation of recombinant chicken cystatin

作　　者：高建梅[1] 杨松[2] 陈冬松[3] 贾丽娇[3] 何金萍[3] 张慧丽[3] 

机构地区：[1]辽宁大学轻型产业学院,辽宁沈阳110036 [2]沈阳军区药品检验所 [3]辽宁大学生命科学院动物资源与疫病防治重点实验室

出　　处：《中国公共卫生》2011年第5期659-660,共2页Chinese Journal of Public Health

基　　金：辽宁省教育厅科技项目(2008226;2008220;LL2010155);辽宁省科学技术厅(20102085)

摘　　要：目的利用具有天然N-末端的重组鸡半胱氨酸蛋白酶抑制剂(chicken cystatin,cC)在毕赤酵母中高效分泌表达,研究淀粉样沉淀倾向。方法通过在巴氏毕赤酵母(Pichia pastoris)菌株X-33中甲醇诱导表达cC能与刚果红特异性结合特性和硫磺素T荧光分析聚积方法研究聚集的特性。结果重组cC形成的类淀粉样蛋白的聚集量有明显增加,30 d时,紫外分光光度检测,重组cC为1.21μmol/L,重组cC荧光强度由1.08增至18.05,即聚集程度增加。结论野生型cystatin C也有形成淀粉样沉淀倾向。Objective To study the tendency of amyloid precipitation of recombinant chicken cysteine proteinase inhibitory（chicken cystatin,cC） with natural N-terminal secreted by yeasts Pichia pastoris.Methods The recombinant cC was expressed in Pasteur yeasts（Pichia pastoris） bacterial strain X-33 with the inducement of the methyl alcohol.The cC could combine specifically with Congo red.Thioflavin T fluorescence was used to detect accumulation characteristics of cC.Results The quantity of amyloid cC accumulation increased significantly.Ultraviolate spectrophotometric detection showed a re-cC content of 1.21μmol/L and fluorescent strength increased from 1.08 to 18.05 with an increased aggregation.Conclusion Wild cystatin C also has the tendency of amyloid precipitation.

关 键 词：重组鸡半胱氨酸蛋白酶抑制剂 天然N-末端 毕赤酵母 类淀粉样特性 二硫键 

分 类 号：Q786[生物学—分子生物学]