Odom O W, Deng H Y, Dabbs E R, Hardesty B
Biochemistry. 1984 Oct 9;23(21):5069-76. doi: 10.1021/bi00316a037.
Escherichia coli ribosomal protein S21 was labeled at its single cysteine group with a fluorescent probe. Labeled S21 showed full activity in supporting MS2 RNA-dependent binding of formylmethionyl-tRNAf to 30S ribosomal subunits. Fluorescence anisotropy measurements and direct analysis on glycerol gradients demonstrate conclusively that labeled S21 binds to 50S ribosomal subunits as well as to 30S and 70S particles. The relative binding affinities are in the order 70S greater than 30S greater than 50S. Other results presented appear to indicate that S21 is bound in the same position on either 50S subunits or 30S subunits as in 70S ribosomes, suggesting that the protein is bound simultaneously to both subunits in the latter. Addition of 50S subunits to 30S particles containing probes on S21 and at the 3' end of 16S RNA caused a decrease in the energy transfer between these points. The results correspond to an apparent change in distance from 51 to 61 A.
用荧光探针标记了大肠杆菌核糖体蛋白S21的单个半胱氨酸基团。标记后的S21在支持甲酰甲硫氨酰 - tRNAf与30S核糖体亚基的MS2 RNA依赖性结合方面表现出完全活性。荧光各向异性测量和对甘油梯度的直接分析确凿地证明,标记后的S21能与50S核糖体亚基以及30S和70S颗粒结合。相对结合亲和力的顺序为70S大于30S大于50S。所呈现的其他结果似乎表明,S21在50S亚基或30S亚基上的结合位置与在70S核糖体中的相同,这表明该蛋白在70S核糖体中同时与两个亚基结合。将50S亚基添加到在S21和16S RNA的3'端含有探针的30S颗粒中,导致这些点之间的能量转移减少。结果对应于距离从51到61埃的明显变化。