A Homo sapiens cytoplasmic ribosomal decoding A-site affinity screen evaluating aminoglycoside and analogue binding.
The potential of aminoglycoside antibiotics to induce premature stop codon read-through in eukaryotic systems has been reported recently, inspiring the evaluation of structural alterations within the Homo sapiens cytoplasmic decoding center on ligand binding. Here we report the employment of an affinity screen capable of monitoring conformational changes of adenines 1492 and 1493 in solution. Thus, changes induced by the presence of a ligand can be directly translated to binding affinities for the eukaryotic decoding center. Binding data for the eukaryotic ribosomal decoding center can be easily obtained by this method and are in excellent agreement with previously reported values measured by alternative techniques. Furthermore, a good correlation is obtained between the experimental binding affinities and the biological activity of the compounds examined. In addition, illustrating the generality of the assay, unnatural rigid aminoglycoside analogues of potential therapeutic significance were evaluated.
Institute of Physical Chemistry, Laboratory of Chemical Biology of Natural Products and Designed Molecules, NCSR Demokritos, 15310 Athens, Greece.
Georgia
G
Kythreoti
Dionisios
D
Vourloumis
D000617:Aminoglycosides / Q000737:chemistry*; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001483:Base Sequence; D006801:Humans; D008024:Ligands; D009690:Nucleic Acid Conformation; D012336:RNA, Ribosomal, 16S / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D008024:Ligands; D012336:RNA, Ribosomal, 16S