Structural characterization of an anti-gp120 RNA aptamer that neutralizes R5 strains of HIV-1

RNA - Tập 11 Số 6 - Trang 873-884 - 2005
Antu Dey1, C. Cohen1, Susan M. Lea1, William James1
1Sir William Dunn School of Pathology and 2Laboratory of Molecular Biophysics, Department of Biochemistry, University of Oxford, Oxford, United Kingdom

Tóm tắt

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-1Ba-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.

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RNA

Tập 11 Số 6

873-884

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