Scanning conformational space with a library of stereo- and regiochemically diverse aminoglycoside derivatives: the discovery of new ligands for RNA hairpin sequences.
A library of stereo- and regiochemically diverse aminoglycoside derivatives was screened at 1 microM using surface plasmon resonance (SPR) against RNA hairpin models of the bacterial A-site, and the HIV viral TAR and RRE sequences. In order to double the stereochemical diversity of the library, the compounds were screened against both enantiomers of each of these sequences. Remarkably, this initial screen suggested that the same four aminoglycoside derivatives bound most tightly to all three of the RNAs, suggesting that these compounds were good RNA binders which, nonetheless, discriminated poorly between the RNA sequences. The interactions between selected isomeric aminoglycoside derivatives and the RNA hairpins were then studied in more detail using an SPR assay. Three isomeric tight-binding aminoglycoside derivatives, which had been identified from the initial screen, were found to bind more tightly to the RNA hairpins (with K(D) values in the range 0.23 to 4.7 microM) than a fourth isomeric derivative (which had K(D) values in the range 6.0 to 30 microM). The magnitude of the tightest RNA-aminoglycoside interactions stemmed, in large part, from remarkably slow dissociation of the aminoglycosides from the RNA targets. The three tight-binding aminoglycoside derivatives were found, however, to discriminate rather poorly between alternative RNA sequences with, at best, around a twenty-fold difference in affinity for alternative RNA hairpin sequences. Within the aminoglycoside derivative library studied, high affinity for an RNA target was not accompanied by good discrimination between alternative RNA sequences.
Ironmonger
Alan
A
School of Chemistry, University of Leeds, Leeds, UK.
Whittaker
Benjamin
B
Baron
Andrew J
AJ
Clique
Blandine
B
Adams
Chris J
CJ
Ashcroft
Alison E
AE
Stockley
Peter G
PG
Nelson
Adam
A
D000617:Aminoglycosides / Q000737:chemistry*; D001483:Base Sequence; D020650:Combinatorial Chemistry Techniques; D016325:HIV Long Terminal Repeat; D008024:Ligands; D008968:Molecular Conformation; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry*; D013237:Stereoisomerism; D020349:Surface Plasmon Resonance
D000617:Aminoglycosides; D008024:Ligands; D012329:RNA, Bacterial; D012367:RNA, Viral