Fluorescence Spectroscopy of tRNAPhe Y Base in the Presence of Mg2+ and Small Molecule Ligands.

• Autores: Todd P. Silverstein, Sarah R. Kirk, Karen McFarlane Holman
• Localización: Journal of chemical education, ISSN 0021-9584, Vol. 85, Nº 5, 2008, págs. 678-679
• Idioma: inglés
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• Resumen

  • This laboratory project is one component of a semester-long advanced biochemistry laboratory course that uses several complementary techniques to study tRNAPhe conformational changes induced by ligand binding. In this article we describe a set of experiments in which students use fluorescence spectroscopy to study tRNAPhe structure. Both intrinsic (Y Base) and extrinsic (ethidium bromide) fluorescence probes are monitored. Ligands (e.g., spermine, neomycin B, magnesium) are titrated into tRNAPhe solution and changes in fluorescence intensity are monitored. Students fit the data to hyperbolic saturation curves to determine ligand-binding equilibrium constants and draw conclusions on how different binding modes affect tRNAPhe tertiary structure.