Fanning T G, Traut R R
Nucleic Acids Res. 1981 Feb 25;9(4):993-1004. doi: 10.1093/nar/9.4.993.
5S RNA-protein complexes were prepared in vitro using partially purified E. coli 5S RNA and total E. coli 70S ribosomal proteins. The complexes were isolated from sucrose gradients and shown to contain proteins L5, L18, L25 and a fourth protein not heretofore characterized and designed L31. The complexes were treated with the crosslinking reagents dimethyl suberimidate and dimethyl-3,3'-dithiobispropionimidate. Both reagents gave identical patterns of crosslinked proteins when analyzed by one-dimensional polyacrylamide/dodecylsulfate gel electrophoresis. Dimers of L5-L31', L5-L18 and L18-L18 and a trimer containing L5, L18 and L31' were identified by diagonal polyacrylamide/dodecylsulfate gel electrophoresis of the proteins crosslinked with dimethyl-3,3'-dithiobispropionimidate. No crosslinking was detected between L25 and the other three proteins.
使用部分纯化的大肠杆菌5S RNA和总的大肠杆菌70S核糖体蛋白在体外制备5S RNA - 蛋白质复合物。这些复合物从蔗糖梯度中分离出来,并显示含有蛋白质L5、L18、L25以及第四种迄今未鉴定的蛋白质,命名为L31。用交联试剂亚胺基二琥珀酸二甲酯和二甲基 - 3,3'-二硫代双丙酰亚胺处理这些复合物。当通过一维聚丙烯酰胺/十二烷基硫酸盐凝胶电泳分析时,两种试剂产生相同的交联蛋白模式。通过对用二甲基 - 3,3'-二硫代双丙酰亚胺交联的蛋白质进行对角线聚丙烯酰胺/十二烷基硫酸盐凝胶电泳,鉴定出L5 - L31'、L5 - L18和L18 - L18的二聚体以及包含L5、L18和L31'的三聚体。未检测到L25与其他三种蛋白质之间的交联。