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Issue 30, 2012
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Probing riboswitch–ligand interactions using thiamine pyrophosphate analogues

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Abstract

The Escherichia coli thiM riboswitch forms specific contacts with its natural ligand, thiamine pyrophosphate (TPP or thiamine diphosphate), allowing it to generate not only nanomolar binding affinity, but also a high degree of discrimination against similar small molecules. A range of synthetic TPP analogues have been used to probe each of the riboswitch–ligand interactions. The results show that the pyrimidine-sensing helix of thiM is exquisitely tuned to select for TPP by recognising the H-bonding donor and acceptors around its aminopyrimidine ring and also by forming π-stacking interactions that may be sensitive to the electronics of the ring. The central thiazolium ring of TPP appears to be more important for ligand recognition than previously thought. It may contribute to binding via long-range electrostatic interactions and/or by exerting an electron withdrawing effect on the pyrimidine ring, allowing its presence to be sensed indirectly and thereby allowing discrimination between thiamine (and its phosphate esters) and other aminopyrimidines found in vivo. The pyrophosphate moiety is essential for submicromolar binding affinity, but unexpectedly, it does not appear to be strictly necessary for modulation of gene expression.

Graphical abstract: Probing riboswitch–ligand interactions using thiamine pyrophosphate analogues

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Publication details

The article was received on 16 Dec 2011, accepted on 08 Mar 2012 and first published on 09 Mar 2012


Article type: Paper
DOI: 10.1039/C2OB07116A
Citation: Org. Biomol. Chem., 2012,10, 5924-5931
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    Probing riboswitch–ligand interactions using thiamine pyrophosphate analogues

    L. Chen, E. Cressina, N. Dixon, K. Erixon, K. Agyei-Owusu, J. Micklefield, A. G. Smith, C. Abell and F. J. Leeper, Org. Biomol. Chem., 2012, 10, 5924
    DOI: 10.1039/C2OB07116A

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