Deng H Y, Odom O W, Hardesty B
Eur J Biochem. 1986 May 2;156(3):497-503. doi: 10.1111/j.1432-1033.1986.tb09608.x.
Isolated Escherichia coli ribosomal protein L11 was labeled with maleimidyl derivatives of coumarin or fluorescein at the thiol group of its single cysteine, then reconstituted singly or in pairs with other fluorescently labeled ribosomal components. The characteristics of fluorescence from the labeled protein were studied and its distance to other components was determined by non-radiative energy transfer. The distance between probes on L11 and cysteine residues on other proteins or the 3' end of the ribosomal RNAs were found to be: S1, 7.4-8.3 nm; S21, 7.6 nm; 23S RNA, 6.9 nm; 5S RNA, 7.6 nm; 16S RNA, greater than 8.5 nm. Considered together with previously published results these distances indicate that the location of L11 in the 50S subunit is below the lateral protuberance characterized by L7/L12.
将分离出的大肠杆菌核糖体蛋白L11在其单个半胱氨酸的巯基处用香豆素或荧光素的马来酰亚胺衍生物进行标记,然后与其他荧光标记的核糖体组分单独或成对重组。研究了标记蛋白的荧光特性,并通过非辐射能量转移确定其与其他组分的距离。发现L11上的探针与其他蛋白质上的半胱氨酸残基或核糖体RNA的3'端之间的距离为:S1,7.4 - 8.3纳米;S21,7.6纳米;23S RNA,6.9纳米;5S RNA,7.6纳米;16S RNA,大于8.5纳米。结合先前发表的结果来看,这些距离表明L11在50S亚基中的位置低于以L7/L12为特征的外侧突起。