Measuring RNA-Ligand Interactions with Microscale Thermophoresis.

Abstract:

:In recent years, there has been dramatic growth in the study of RNA. RNA has gone from being known as an intermediate in the central dogma of molecular biology to a molecule with a large diversity of structure and function that is involved in all aspects of biology. As new functions are rapidly discovered, it has become clear that there is a need for RNA-targeting small molecule probes to investigate RNA biology and clarify the potential for therapeutics based on RNA-small molecule interactions. While a host of techniques exist to measure RNA-small molecule interactions, many of these have drawbacks that make them intractable for routine use and are often not broadly applicable. A newer technology called microscale thermophoresis (MST), which measures the directed migration of a molecule and/or molecule-ligand complex along a temperature gradient, can be used to measure binding affinities using very small amounts of sample. The high sensitivity of this technique enables measurement of affinity constants in the nanomolar and micromolar range. Here, we demonstrate how MST can be used to study a range of biologically relevant RNA interactions, including peptide-RNA interactions, RNA-small molecule interactions, and displacement of an RNA-bound peptide by a small molecule.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Moon MH,Hilimire TA,Sanders AM,Schneekloth JS Jr

doi

10.1021/acs.biochem.7b01141

subject

Has Abstract

pub_date

2018-08-07 00:00:00

pages

4638-4643

issue

31

eissn

0006-2960

issn

1520-4995

journal_volume

57

pub_type

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