| Autor: |
Wendeler, Michaela, T. Miller, Jennifer, S. Baichoo, Noel, Le Grice, Stuart, Lee, Hsiu-Fang, Kvaratskhelia, Mamuka, Bermingham, Alun, Beutler, John, R. O’Keefe, Barry, Chertov, Oleg, K. Bona, Marion, K. Bona, Marion, Ehteshami, Maryam, Götte, Matthias |
| Zdroj: |
ACS Chemical Biology; October 2008, Vol. 3 Issue: 10 p635-644, 10p |
| Abstrakt: |
High-throughput screening of National Cancer Institute libraries of synthetic and natural compounds identified the vinylogous ureas 2-amino-5,6,7,8-tetrahydro-4 H-cyclohepta[ b]thiophene-3-carboxamide (NSC727447) and N-[3-(aminocarbonyl)-4,5-dimethyl-2-thienyl]-2-furancarboxamide (NSC727448) as inhibitors of the ribonuclease H (RNase H) activity of HIV-1 and HIV-2 reverse transcriptase (RT). A Yonetani−Theorell analysis demonstrated that NSC727447, and the active-site hydroxytropolone RNase H inhibitor β-thujaplicinol were mutually exclusive in their interaction with the RNase H domain. Mass spectrometric protein footprinting of the NSC727447 binding site indicated that residues Cys280 and Lys281 in helix I of the thumb subdomain of p51 were affected by ligand binding. Although DNA polymerase and pyrophosphorolysis activities of HIV-1 RT were less sensitive to inhibition by NSC727447, protein footprinting indicated that NSC727447 occupied the equivalent region of the p66 thumb. Site-directed mutagenesis using reconstituted p66/p51 heterodimers substituted with natural or non-natural amino acids indicates that altering the p66 RNase H primer grip significantly affects inhibitor sensitivity. NSC727447 thus represents a novel class of RNase H antagonists with a mechanism of action differing from active site, divalent metal-chelating inhibitors that have been reported. |
| Databáze: |
Supplemental Index |
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