UV Thermal Melting Curves of tRNAPhe in the Presence of Ligands

摘要

This laboratory project is one component of a semester-long advanced biochemistry laboratory course that uses several complementary techniques to study tRNAPhe conformational chvanges induced by ligand binding. In this article we describe a set of experiments in which the thermal unfolding of tRNAPhe is studied with UV–vis spectrophotometry. Over the course of two laboratory periods, students examine the effects of magnesium and small molecule ligands on tRNAPhe conformational stability. The data collected in this laboratory are fit to saturation binding curves, the ligand-binding equilibrium constants are determined, and conclusions are drawn about tRNAPhe ligand binding modes and the impact of various types of ligands on tRNA structure.