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Involvement of the 3' side of the anti codon loop of yeast tyrosine transfer rna in messenger free binding to ribosomes an esr study

, : Involvement of the 3' side of the anti codon loop of yeast tyrosine transfer rna in messenger free binding to ribosomes an esr study. European Journal of Biochemistry 139(3): 541-546

ESR spectra of a nitroxide spin-label attached to residue i6A-37 of yeast tRNATyr were measured in complexes of deacylated tRNATyr with Escherichia coli ribosomes. A Scatchard plot, obtained in the absence of mRNA, indicated strong binding with an association constant of 1 .times. 107 l .cntdot. mol-1, suggesting the P-site binding. The ESR spectrum of free tRNATyr, characteristic for a rapidly tumbling nitroxide, changes to a spectrum with extensively broadened lines in the ribosome-tRNA complex. The original spectrum can be restored upon long incubations of the complex with an excess of extraneous tRNA. ESR spectra suggest that the spin-label motion is drastically perturbed though not completely blocked in the ribosome-tRNATyr complex. The ESR spectra of a spin-label attached to the opposite, i.e., 5', side of the anticodon loop are only slightly perturbed by the messenger-free binding to ribosomes, indicating that the 2 sides of the anticodon loop face entirely different environments when bound to the P site, the 3' side being oriented towards the surface of the ribosome, and the other side towards its environment or a large cavity.


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