Vinylogous ureas as a novel class of inhibitors of reverse transcriptase-associated ribonuclease H activity
| Autor: | Oleg Chertov, Hsiu-Fang Lee, Maryam Ehteshami, Mamuka Kvaratskhelia, John A. Beutler, Matthias Götte, Alun Bermingham, Jennifer T. Miller, Barry R. O'Keefe, Michaela Wendeler, Noel S. Baichoo, Stuart F. J. Le Grice, Marion K. Bona |
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| Rok vydání: | 2008 |
| Předmět: |
DNA polymerase
Thiophenes Biochemistry Article Combinatorial Chemistry Techniques Humans Urea Binding site RNase H Furans chemistry.chemical_classification Binding Sites biology Protein footprinting Active site General Medicine Reverse transcriptase HIV Reverse Transcriptase Amino acid Ribonuclease H Human Immunodeficiency Virus chemistry biology.protein Molecular Medicine Reverse Transcriptase Inhibitors Primer (molecular biology) |
| Zdroj: | ACS chemical biology. 3(10) |
| ISSN: | 1554-8937 |
| Popis: | 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 beta-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: | OpenAIRE |
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