A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35?NP Interface [electronic resource]

• Ngôn ngữ: eng
• Ký hiệu phân loại: 572.8 Biochemical genetics
• Thông tin xuất bản: Bethesda, Md. : Oak Ridge, Tenn. : National Institutes of Health (U.S.) ; Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy, 2017
• Mô tả vật lý: Size: p. 190-198 : , digital, PDF file.
• Bộ sưu tập: Metadata
• ID: 260654
• Chủ đề: 1. 60 applied life sciences; 2. Assays; 3. Dna replication; 4. Ebola virus; 5. Et al; 6. Fluorescence; 7. Fluorescence polarization assay; 8. High-throughput screening; 9. Nucleoprotein; 10. Peptides and proteins; 11. Vp35

Tóm tắt

The 2014 Ebola outbreak in West Africa, the largest outbreak on record, highlighted the need for novel approaches to therapeutics targeting Ebola virus (EBOV). Within the EBOV replication complex, the interaction between polymerase cofactor, viral protein 35 (VP35), and nucleoprotein (NP) is critical for viral RNA synthesis. Here, we recently identified a peptide at the N-terminus of VP35 (termed NPBP) that is sufficient for interaction with NP and suppresses EBOV replication, suggesting that the NPBP binding pocket can serve as a potential drug target. Here we describe the development and validation of a sensitive high-throughput screen (HTS) using a fluorescence polarization assay. Initial hits from this HTS include the FDA-approved compound tolcapone, whose potency against EBOV infection was validated in a nonfluorescent secondary assay. High conservation of the NP?VP35 interface among filoviruses suggests that this assay has the capacity to identify pan-filoviral inhibitors for development as antivirals.