Structural characterization of an anti-gp120 RNA aptamer that neutralizes R5 strains of HIV-1.
We recently described the isolation of 2'-fluoropyrimidine-substituted RNA aptamers that bind specifically to the surface glycoprotein (gp 120) of the R5 strain, HIV-1(Ba-L), as presented in a previous study. These aptamers potently neutralize HIV-1 infectivity in human peripheral blood mononuclear cells of both tissue culture lab-adapted strains and diverse R5 clinical isolates from multiple clades. Here, we report a detailed structural characterization of one such neutralizing aptamer, B40, using enzymatic and chemical probing methods. We identify the minimal region of the aptamer essential for binding gp120 and accordingly design a 77-nucleotide truncated aptamer, B40t77. We then quantitatively analyze the binding affinity and neutralization potency of the parental and truncated (minimal) aptamer, and show them to be comparable. Furthermore, using results from secondary structure analysis, RNA mutagenesis and BIAcore surface plasmon resonance (SPR) binding assays, we hypothesize that a folded RNA structure is required to present specific nucleotide sequences to allow gp120-recognition and binding. The information gained from this study may provide leads for development of novel anti-HIV-1 therapies and can be used to extend our understanding of the molecular interactions between the virus and its host cell.
Dey
Antu K
AK
Sir William Dunn School of Pathology, University of Oxford, UK.
Griffiths
Carla
C
Lea
Susan M
SM
James
William
W
D000818:Animals; D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology; D052157:Aptamers, Nucleotide; D001483:Base Sequence; D002478:Cells, Cultured; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D009154:Mutation; D009690:Nucleic Acid Conformation; D011743:Pyrimidines / Q000737:chemistry; D012313:RNA / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology
D019380:Anti-HIV Agents; D052157:Aptamers, Nucleotide; D015699:HIV Envelope Protein gp120; D011743:Pyrimidines; C502124:RNA aptamer B40; D012313:RNA