The use of a new series of cleavable protein‐crosslinkers on the Escherichia coli ribosome
The use of a new series of cleavable protein‐crosslinkers on the Escherichia coli ribosome
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一系列新的可裂解蛋白质交联剂在大肠杆菌核糖体上的使用
DOI：
10.1016/0014-5793(74)80488-6
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发表时间：
1974
期刊：
影响因子：
3.5
通讯作者：
L. Lutter
中科院分区：
文献类型：
--
作者：
L. Lutter
蛋白质-蛋白质交联试剂已被广泛用于研究大肠杆菌核糖体中的蛋白质排列[l-9]，但这些研究一直受到鉴定新形成的交联复合物成分的困难的阻碍。一种可以在温和条件下裂解的交联试剂将极大地促进这一鉴定步骤。含有二硫化物桥的交联试剂已经被描述过[9,10]，但这些交联试剂在分离交联配合物的过程中排除了还原剂的使用，因此，可以想象，非邻近的蛋白质可以通过溶液中的二硫化物交换连接在一起。在本研究中，我们描述了一系列含有邻羟基的不同桥长化学交联试剂。这些试剂形成的交联可以通过高碘酸盐的温和处理来定量地断裂。还描述了两种聚丙烯酰胺凝胶电泳系统，其有利于形成的配合物的分析。
Protein-protein crosslinking reagents have been used extensively to study the protein arrangements in the Escherichia coli ribosome [l-9], but these studies have been hindered by the difficulty of identifying the constituents of the new crosslinked complexes formed. A crosslinking reagent which could be cleaved under mild conditions would greatly facilitate this identification step. Crosslinking reagents containing a disulfide bridge have been described [9, 10], but these preclude the use of reducing agents during the isolation of crosslinked complexes and, therefore, could conceivably allow non-neighboring proteins to become linked together through disulfide interchange in solution. In the present study we describe a new series of chemical crosslinking reagents of varying bridge length which contain vicinal hydroxyl groups. The crosslinks formed by these reagents can be quantitatively cleaved by mild treatment with periodate. Two polyacrylamide gel electrophoresis systems are also described which facilitate analysis of the complexes formed.