Development and Validation of a High-Throughput Screening Assay for the Hepatitis C Virus p7 Viroporin

Gervais, Christian; Dô, Florence; Cantin, Ariane; Kukolj, George; White, Peter W.; Gauthier, Annick; Vaillancourt, Frédéric H.

doi:10.1177/1087057110396215

Cited by 21 publications

(20 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As previously suggested for HCV p7 (Gervais et al, 2011;StGelais et al, 2007), reconstitution of CSFV p7 viroporin function in a liposomal system may be useful for identifying and characterizing new potential antiviral compounds. However, the poor yield of synthesis, owing to the length and hydrophobicity of the sequence, restricts the potential application of the complete protein to systematic experimentation.…”

Section: Discussionmentioning

confidence: 95%

Pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function

et al. 2014

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 95%

Pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function

et al. 2014

View full text Add to dashboard Cite

“…33 In many cases, however, the successful use of liposomes depends on their formulation and the method of preparation and can be used for robust high-throughput screening assays. 34 …”

Section: Introductionmentioning

confidence: 99%

Role of Nanoparticle Surface Functionality in the Disruption of Model Cell Membranes

et al. 2012

View full text Add to dashboard Cite

Lipid bilayers are biomembranes common to cellular life and constitute a continuous barrier between cells and their environment. Understanding the interaction of engineered nanomaterials (ENMs) with lipid bilayers is an important step toward predicting subsequent biological effects. In this study, we assess the effect of varying the surface functionality and concentration of 10 nm-diameter gold (Au) and titanium dioxide (TiO2) ENMs on the disruption of negatively charged lipid bilayer vesicles (liposomes) using a dye leakage assay. Our findings show that Au ENMs having both positive and negative surface charge induce leakage that reaches a steady state after several hours. Positively charged particles with identical surface functionality and different core composition show similar leakage effects and result in faster and greater leakage than negatively charged particles, which suggests that surface functionality, not particle core composition, is a critical factor in determining the interaction between ENMs and lipid bilayers. The results suggest that particles permanently adsorb to bilayers and that only one positively charged particle is required to disrupt a liposome and trigger leakage of its entire contents in contrast to mellitin molecules, the most widely studied membrane lytic peptide, which requires hundred of molecules to generate leakage.

show abstract

“…Screening based upon liposome dyerelease assays conducted by Boehringer Ingelheim was found to be robust, generating few false-positives and a sensible percentage hit rate, although this has not been followed up to date in the literature (Gervais et al, 2011). Moreover, rational compound design based upon the adamantane binding site in PDB ID: 3ZD0 structure-guided channel models yielded compounds with much improved potency, with nanomolar IC 50 values against HCV in culture (Foster et al, 2014).…”

Section: Hcv P7mentioning

confidence: 99%

“…This was exemplified by the use of liposome dye-release assays to conduct a high-throughput screen of potential HCV p7 small-molecule inhibitors (Gervais et al, 2011). Indeed, many viroporins with variable structures and functions have been shown to conduct diverse small molecules in addition to ionic species.…”

Section: General Viroporin Characteristicsmentioning

confidence: 99%

Viroporins: structure, function and potential as antiviral targets

Scott

Griffin

2015

Journal of General Virology

121

146

View full text Add to dashboard Cite

The channel-forming activity of a family of small, hydrophobic integral membrane proteins termed 'viroporins' is essential to the life cycles of an increasingly diverse range of RNA and DNA viruses, generating significant interest in targeting these proteins for antiviral development. Viroporins vary greatly in terms of their atomic structure and can perform multiple functions during the virus life cycle, including those distinct from their role as oligomeric membrane channels. Recent progress has seen an explosion in both the identification and understanding of many such proteins encoded by highly significant pathogens, yet the prototypic M2 proton channel of influenza A virus remains the only example of a viroporin with provenance as an antiviral drug target. This review attempts to summarize our current understanding of the channel-forming functions for key members of this growing family, including recent progress in structural studies and drug discovery research, as well as novel insights into the life cycles of many viruses revealed by a requirement for viroporin activity. Ultimately, given the successes of drugs targeting ion channels in other areas of medicine, unlocking the therapeutic potential of viroporins represents a valuable goal for many of the most significant viral challenges to human and animal health.

show abstract

Development and Validation of a High-Throughput Screening Assay for the Hepatitis C Virus p7 Viroporin

Cited by 21 publications

References 15 publications

Pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function

Pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function

Role of Nanoparticle Surface Functionality in the Disruption of Model Cell Membranes

Viroporins: structure, function and potential as antiviral targets

Contact Info

Product

Resources

About