Sihag R K, Deutscher M P
J Biol Chem. 1983 Oct 10;258(19):11846-50.
In order to gain some insight into the structural parameters important for aminoacyl-tRNA synthetase complex formation, we have examined the effect of various salts and detergents on the stability and structure of the synthetase complex. Certain neutral salts were found to inactivate aminoacyl-tRNA synthetase activities in the complex, and the order of effectiveness in this process followed a classical Hofmeister series. In addition, one of these salts, NaSCN, was also effective in partially dissociating the complex. Detergents varied in their ability to inactivate synthetases, with ionic detergents being most effective and nonionic detergents being much less destructive. Detergents, by themselves, could partially disrupt the complex; however, in the presence of 1 M NaCl, nonionic detergents did lead to considerable dissociation of synthetases and generation of low molecular weight forms of these enzymes. Removal of lipids from the complex with the nonionic detergent, Triton X-114, rendered arginyl-tRNA synthetase sensitive to the addition of NaCl. However, this salt sensitivity was abolished by readdition of a lipid extract isolated from the complex. These results implicate hydrophobic interactions in the stability of the synthetase complex, and suggest the possible involvement of lipids in maintaining its structural integrity.
为了深入了解对氨酰 - tRNA合成酶复合物形成至关重要的结构参数,我们研究了各种盐和去污剂对合成酶复合物稳定性和结构的影响。发现某些中性盐会使复合物中的氨酰 - tRNA合成酶活性失活,并且在此过程中的有效性顺序遵循经典的霍夫迈斯特序列。此外,其中一种盐硫氰酸钠(NaSCN)在部分解离复合物方面也很有效。去污剂使合成酶失活的能力各不相同,离子型去污剂最有效,而非离子型去污剂的破坏性要小得多。单独使用去污剂可部分破坏复合物;然而,在1 M NaCl存在下,非离子型去污剂确实导致合成酶大量解离并产生这些酶的低分子量形式。用非离子型去污剂Triton X - 114从复合物中去除脂质后,精氨酰 - tRNA合成酶对添加NaCl变得敏感。然而,重新添加从复合物中分离出的脂质提取物可消除这种盐敏感性。这些结果表明疏水相互作用对合成酶复合物的稳定性有影响,并表明脂质可能参与维持其结构完整性。