Direct observation of aminoglycoside-RNA binding by localized surface plasmon resonance spectroscopy.
RNA is involved in fundamental biological functions when bacterial pathogens replicate. Identifying and studying small molecules that can interact with bacterial RNA and interrupt cellular activities is a promising path for drug design. Aminoglycoside (AMG) antibiotics, prominent natural products that recognize RNA specifically, exert their biological functions by binding to prokaryotic ribosomal RNA and interfering with protein translation, ultimately resulting in bacterial cell death. The decoding site, a small internal loop within the 16S rRNA, is the molecular target for the AMG antibiotics. The specificity of neomycin B, a highly potent AMG antibiotic, to the ribosomal decoding RNA site, was previously studied by observing AMG-RNA complexes in solution. Here, we study this interaction using localized surface plasmon resonance (LSPR) transducers comprising gold island films prepared by evaporation on glass and annealing. Small molecule AMG receptors were immobilized on the Au islands via polyethylene glycol (PEG)-thiol linkers, and the interaction with target RNA in solution was studied by monitoring the change in the LSPR optical response upon binding. The results show high-affinity binding of neomycin to 27-nucleotide model A-site RNA sequence in the nanomolar range, while no specific binding is observed for synthetic RNA oligomers (e.g., poly-U). The impact of specific base substitutions in the A-site RNA constructs on binding affinity and selectivity is determined quantitatively. It is concluded that LSPR is a powerful tool for providing molecular insight into small molecule-RNA interactions and for the design and screening of selective antimicrobial drugs.
Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot 76100, Israel.
Ludmila
L
Frolov
Andrew
A
Dix
Yitzhak
Y
Tor
Alexander B
AB
Tesler
Yulia
Y
Chaikin
Alexander
A
Vaskevich
Israel
I
Rubinstein
D000617:Aminoglycosides / Q000737:chemistry / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D005601:Framycetin / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D007700:Kinetics; D053768:Metal Nanoparticles / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000378:metabolism*; D055672:Static Electricity; D020349:Surface Plasmon Resonance
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D012336:RNA, Ribosomal, 16S; D011092:Polyethylene Glycols; D005601:Framycetin; D006046:Gold