2-aminopurine fluorescence: discrimination between specific and unspecific ligand binding to the kissing-loop dimer of the HIV-1 RNA.
The fluorescent 2-aminopurine probe (2-AP) incorporated into the loop of 23-mer RNA hairpin of HIV-1 genome dimerization initiation site (DIS) was used for discrimination of specific and unspecific binding of paromomycin and spermine to the kissing loop dimer (KD) formed in solution. While both ligands stabilized the KD RNA structure, only paromomycin binding resulted in significant increase of 2-AP fluorescence. These observations suggest that the 2-AP fluorescent RNA construct might be useful for selecting ligands specifically binding the HIV-1 kissing loop RNA dimer.
Kaluzhny
Dmitry N
DN
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilova, 32, Moscow, 199911, Russia. dkaluzhny@gmail.com
Beniaminov
Artemy D
AD
Minyat
Elvira E
EE
D015075:2-Aminopurine / Q000737:chemistry*; D019380:Anti-HIV Agents / Q000737:chemistry*; D001665:Binding Sites; D019281:Dimerization; D024202:Electrophoretic Mobility Shift Assay; D005456:Fluorescent Dyes / Q000737:chemistry*; D015497:HIV-1 / Q000235:genetics*; D008024:Ligands; D010303:Paromomycin / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry*; D013096:Spermine / Q000737:chemistry
D019380:Anti-HIV Agents; D005456:Fluorescent Dyes; D008024:Ligands; D012367:RNA, Viral; D013096:Spermine; D015075:2-Aminopurine; D010303:Paromomycin