Universal isolation of cross-linked peptides: application to neurofibrillary tangles.
Universal isolation of cross-linked peptides: application to neurofibrillary tangles.
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交联肽的通用分离：在神经原纤维缠结中的应用。
DOI：
10.1021/bc980098v
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Anderson,VE
中科院分区：
文献类型：
--
作者：
Chen,X;Eswaran,D;Smith,MA;Perry,G;Anderson,VE
已经证明了分离交联肽的通用程序。该程序依赖于最初通过用氰基硼氢化钠和甲醛还原甲基化将赖氨酸的ε-氨基转化为二甲基赖氨酸。赖氨酸修饰的蛋白质被蛋白水解裂解，所得的α-氨基基团用甲氧基聚（乙二醇）5000琥珀酰羟基琥珀酰亚胺衍生。酪氨酸侧链的任何无意衍生化可通过在弱碱性条件下孵育来逆转。交联的多肽含有两个聚（乙二醇）5000链，而非交联的肽含有一个单一的聚（乙二醇）5000链。这两种肽群可以通过凝胶过滤色谱法分离。该方法已被证明能够使用谷胱甘肽、溶菌酶、化学交联血红蛋白和从阿尔茨海默病病例的脑中分离的神经元缠结来分离交联肽。
A universal procedure to isolate cross-linked peptides has been demonstrated. The procedure relies on initially converting the ε-amino groups of lysine to dimethyl lysine by reductive methylation with sodium cyanoborohydride and formaldehyde. The lysine-modified protein is proteolytically cleaved and the resulting α-amino groups derivatized with methoxypoly(ethylene glycol)5000succinyl hydroxysuccinimide. Any unintentional derivatization of tyrosine side chains can be reversed by incubation under mildly alkaline conditions. The cross-linked polypeptides contain two poly(ethylene glycol)5000chains while non-cross-linked peptides contain a single poly(ethylene glycol)5000chain. The two populations of peptides can be separated by gel filtration chromatography. This procedure has been shown capable of isolating cross-linked peptides using glutathione, lysozyme, chemically cross-linked hemoglobin, and neurofibrillary tangles isolated from the brain of a case of Alzheimer's disease.