Separation and analysis of the soluble trimer of Aβ_(1-40) and its effects on the rise in intracellular calcium  被引量:1

Separation and analysis of the soluble trimer of Aβ_(1-40) and its effects on the rise in intracellular calcium

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作  者:ZHENG Xiaohui WANG Lijun ZHANG Lan HONG Yuankai HUANG Lixin SHA Yinlin 

机构地区:[1]Single-molecule and Nanobiology Laboratory. Department of Biophysics School of Basic Medical Sciences, Peking University, Beijing 100083, China

出  处:《Chinese Science Bulletin》2006年第7期830-838,共9页

基  金:supported by the National Natural Science Foundation of China(Grant Nos.20273002&20103001).

摘  要:The oligomers of Aβ1-40 peptide in PBS buffer solution were analyzed by SEC and native PAGE, and the trimer of Aβ1-40 was also isolated by SEC. In addition, the effects of the soluble Aβ1-40 trimer on intracellular free calcium (Ca2+) balance of hippocampal neurons of postnatal rats were investi- gated by fluorescence microscopy. The experimental results indicated that Aβ1-40 peptide existed in the form of low molecular weight oligomers in 0.231 mmol/L fresh Aβ1-40 solution (20 mmol/L sodium phosphate buffer, pH 7.4, 0.02% sodium azide) within 24 h and the soluble trimer was the most abundant species. Both the trimeric and the fibrillar Aβ1-40 were able to increase the intracellular Ca2+ concentration, but the Aβ1-40 trimer caused a gradual rise and the potential was also stronger than that of the fibrils at the same concentration. In addition there were dif- ferent response modes for trimeric and fibrillar Aβ1-40, meaning that there are different mechanisms of in- crease in intracellular Ca2+ caused by Aβ1-40.The oligomers of Aβ(1-40)peptide in PBS buffer solution were analyzed by SEC and native PAGE, and the trimer of Aβ(1-40) was also isolated by SEC. In addition, the effects of the soluble Aβ(1-40) trimer on intracellular free calcium (Ca^2+) balance of hippocampal neurons of postnatal rats were investigated by fluorescence microscopy. The experimental results indicated that Aβ(1-40) peptide existed in the form of low molecular weight oligomers in 0.231 mmol/L fresh Aβ(1-40) solution (20 mmol/L sodium phosphate buffer, pH 7.4, 0.02% sodium azide) within 24 h and the soluble trimer was the most abundant species. Both the trimeric and the fibrillar Aβ(1-40) were able to increase the intracellular Ca^2+ concentration, but the Aβ(1-40) trimer caused a gradual rise and the potential was also stronger than that of the fibrils at the same concentration. In addition there were different response modes for trimeric and fibrillar Aβ(1-40). meaning that there are different mechanisms of increase in intracellular Ca^2+ caused by Aβ(1-40).

关 键 词: 三聚物 分离 SEC 色谱分析 

分 类 号:R96[医药卫生—药理学]

 

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