Binding of S21 to the 50S subunit and the effect of the 50S subunit on nonradiative energy transfer between the 3' end of 16S RNA and S21

• Published in: Biochemistry (Easton) Vol. 23; no. 21; pp. 5069 - 5076
• Main Authors: Odom, Obed W, Deng, Hung Yin, Dabbs, Eric R, Hardesty, Boyd
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 09.10.1984
• Subjects: Biological and medical sciences; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fluorescence Polarization; Fundamental and applied biological sciences. Psychology; Interactions. Associations; Intermolecular phenomena; Macromolecular Substances; Molecular biophysics; Molecular Weight; Mutation; Peptide Initiation Factors - metabolism; Ribosomal Proteins - metabolism; Ribosomes - metabolism; RNA, Ribosomal - metabolism; RNA, Transfer, Amino Acyl - metabolism; Molecular arrangement; Fluorescent labelling; Escherichia coli; 50S-Ribosome; 30S-Ribosome; 16S-RNA; Bacteria; Molecular interaction; Excitation energy transfer; Escherichieae; Enterobacteriaceae
• ISSN: 0006-2960; 1520-4995
• 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.