A Time-Resolved Fluorescence–Resonance Energy Transfer Assay for Identifying Inhibitors of Hepatitis C Virus Core Dimerization

Kota, Smitha; Scampavia, Louis; Spicer, Timothy; Beeler, Aaron B.; Takahashi, Virginia; Snyder, John K.; Porco, John A.; Hodder, Peter; Strosberg, A. Donny

doi:10.1089/adt.2009.0217

Cited by 25 publications

(52 citation statements)

References 24 publications

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“…1D). In comparison, ebselen presented no activity on HCV core C106 dimerization by TR-FRET assays (23).…”

Section: Hts-tr-fret For the Identification Of Inhibitors Of Ctd Dimementioning

confidence: 89%

“…The TR-FRET signal was calculated as a ratio of emission: (665 nm/620 nm) ϫ 10,000. Hepatitis C virus (HCV) core protein was used as a counterscreen (23). All plates were analyzed and passed quality control (QC) if their Z= score was greater than 0.5.…”

Section: Tr-fretmentioning

confidence: 99%

“…The top 10 hit compounds inhibited CTD-CTD interaction by 59% to 71%. The LOPAC library was cross-referenced for HCV core inhibitors (23), and out of the 42 hits identified in our screen, 9 displayed inhibitory activity against HCV core dimerization (15.7% to 51.6%). We tested 20 of the top hits in multiple rounds of HIV-1 infection in HeLa-CD4 cells and identified ebselen, an organoselenium compound (Fig.…”

Section: Hts-tr-fret For the Identification Of Inhibitors Of Ctd Dimementioning

confidence: 99%

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Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication

Thenin-Houssier

Vera

Pedró-Rosa

et al. 2016

Antimicrob Agents Chemother

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102

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“…1D). In comparison, ebselen presented no activity on HCV core C106 dimerization by TR-FRET assays (23).…”

Section: Hts-tr-fret For the Identification Of Inhibitors Of Ctd Dimementioning

confidence: 89%

Section: Tr-fretmentioning

confidence: 99%

Section: Hts-tr-fret For the Identification Of Inhibitors Of Ctd Dimementioning

confidence: 99%

See 1 more Smart Citation

Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication

Thenin-Houssier

Vera

Pedró-Rosa

et al. 2016

Antimicrob Agents Chemother

Self Cite

102

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“…The ir use a s fl uorophores in F RET has b een more limited to date, a nd may b e due in part to t heir relatively large size -although some of th e smaller materials being develop ed (e .g., Per CP and CryptoFluor TM ) may address this c onc ern. That said, th ey have be en demonst rated i n sandwic h-based p eptid e and a ntibody FRET a ssays for target analyte detect ion and f or de mons trating FRET in combination with Au a nd QD NMs [121,257,[260][261][262][263][264]. The combination of APC (acceptor) and Eu-cryp t ate (donor ) h as b een use d for th e time-res olve d FRET-based detection of telomerase activity [262], inhibitors of hepatitis C virus core dimerization [264], and small-molecule inhibitors of HIV-1 fusion [263].…”

Section: Green Fluorescent Protein and Derivativesmentioning

confidence: 99%

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

Sapsford

Wildt

Mariani

et al. 2013

FRET – Förster Resonance Energy Transfer

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The intrinsic sensitivity (r 6 dependence) of F€ orster (or fluorescence) resonance energy transfer (FRET) to nanoscale changes in the donor/acceptor separation distance has made FRET an invaluable biophysical tool in a variety of applications, ranging from studying the structure and conformation of proteins and nucleic acids to examining biomolecular interactions, including its use in in vitro and in vivo bioassays [1][2][3][4][5][6][7]. While the myriad of FRET configurations and techniques currently in use are covered throughout this book, here we focus primarily on the materials utilized as donor or acceptor probes in FRET rather than the process itself [3,5,8,9]. Our 2006 review paper on this topic serves as the foundation for this updated chapter [3]. The materials were divided into three main categories: organic materials that include "traditional" dye fluorophores, dark quenchers, polymers, and carbon nanomaterials (NMs); inorganic materials such as metal chelates, metal, and semiconductor nanocrystals; and fluorophores of biological origin such as fluorescent proteins (FPs), amino acids, and fluorescence generated from enzymatic bioluminescence (BL) and chemiluminescence (CL). These materials may function as FRET donors and/or acceptors, depending upon experimental design. Many of the new materials developed and/or new donor-acceptor probe combinations used address some of the inherent complications of more traditional FRET materials, including photobleaching, spectral cross talk, and direct excitation of the acceptor species, and examples of these will be highlighted and discussed throughout the chapter. Since the vast majority of FRET applications are biological in nature, they routinely involve some type of biomolecule labeling strategy, which ultimately plays a significant and fundamental role in the success and interpretation of the resulting FRET. Therefore, we begin the chapter with a brief discussion of the bioconjugation techniques commonly utilized for FRET and points to consider, followed by sections highlighting the current and emerging materials that are used, or have the potential to be used, in FRET applications.

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“…Approaches to develop antiviral molecules targeting viral proteins are only starting to emerge. A screening test for molecules inhibiting core dimerisation has indeed recently been developed 147. The p7 polypeptide is another potential target, however the currently available compounds against this protein have a modest antiviral effect (reviewed in Steinmann and Pietschmann148).…”

Section: Hcv Assembly As a Therapeutic Targetmentioning

confidence: 99%

New advances in the molecular biology of hepatitis C virus infection: towards the identification of new treatment targets

Ploß

Dubuisson

2012

Gut

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Hepatitis C virus (HCV) causes chronic infection in almost 2% of the world's population. If untreated, chronic carriers can develop severe liver disease including fibrosis, cirrhosis and hepatocellular carcinoma. Until recently, hepatitis C was treated with a combination of pegylated interferon and ribavirin, a treatment which was only partially effective and was plagued with side effects. In 2011 two inhibitors of the virally encoded NS3/4 protease have become part of standard therapy, which have improved treatment rates but can exacerbate the problematic side effects. While the addition of these first directly acting antivirals (DAAs) marks a milestone in anti-HCV therapy, new and improved combinations of drugs are desperately needed. New generations of drugs will have to address genetic variability of HCV and issues of viral resistance. Furthermore, combination therapies have to be tailored to effectively cure patient populations that have traditionally been hardest to treat, including patients with cirrhosis, those receiving liver transplants and individuals who are co-infected with HIV or hepatitis B virus. Since the discovery of HCV a plethora of experimental tools have been developed which enabled detailed analysis of various aspects of the viral life cycle and the interaction of HCV with its human host. Such studies have revealed a growing list of targets for therapeutic intervention, some of which will be discussed in this review.

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A Time-Resolved Fluorescence–Resonance Energy Transfer Assay for Identifying Inhibitors of Hepatitis C Virus Core Dimerization

Cited by 25 publications

References 24 publications

Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication

Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

New advances in the molecular biology of hepatitis C virus infection: towards the identification of new treatment targets

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