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BioAssay: AID 493079

Late stage assay provider results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Fluorescence polarization-based biochemical dose response assay for EBNA-1 inhibitors

Name: Late stage assay provider results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Fluorescence polarization-based biochemical dose response assay for EBNA-1 inhibitors. ..more
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 Tested Compounds
 Tested Compounds
All(30)
 
 
Active(1)
 
 
Inactive(29)
 
 
 Tested Substances
 Tested Substances
All(30)
 
 
Active(1)
 
 
Inactive(29)
 
 
AID: 493079
Data Source: The Scripps Research Institute Molecular Screening Center (EBNA1_INH_FP_IC50 MDRUN Assay Provider)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network, Assay Provider
BioAssay Version:
Deposit Date: 2011-01-24
Hold-until Date: 2012-01-21
Modify Date: 2012-01-24

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compound: 1
Related Experiments
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AIDNameTypeComment
1950Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1).Screeningdepositor-specified cross reference: Primary screen (EBNA-1 inhibitors in singlicate)
1955Summary of probe development efforts to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1).Summarydepositor-specified cross reference: Summary (EBNA-1 inhibitors)
2234Counterscreen for inhibitors of EBNA-1: fluorescence polarization-based biochemical high throughput primary assay to identify inhibitors of the Epstein-Barr virus-encoded protein, ZTA.Screeningdepositor-specified cross reference: Counterscreen (ZTA inhibitors in singlicate)
2292Fluorescence polarization-based biochemical high throughput confirmation assay to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1).Screeningdepositor-specified cross reference: Confirmation screen (EBNA-1 inhibitors in triplicate)
2328Counterscreen for inhibitors of EBNA-1: fluorescence polarization-based biochemical high throughput assay for inhibitors of the Epstein-Barr virus-encoded protein, ZTA, in triplicate.Screeningdepositor-specified cross reference: Counterscreen (ZTA inhibitors in triplicate)
2377Counterscreen for inhibitors of EBNA-1: fluorescence polarization-based biochemical high throughput dose response assay to identify inhibitors of the Epstein-Barr virus-encoded protein, ZTAConfirmatorydepositor-specified cross reference: Dose response counterscreen (ZTA inhibitors in triplicate)
2381Fluorescence polarization-based biochemical high throughput dose response assay for inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1)Confirmatorydepositor-specified cross reference: Dose response (EBNA-1 inhibitors in triplicate)
588498qHTS profiling assay for firefly luciferase inhibitor/activator using purifed enzyme and Km concentrations of substrates (counterscreen for the Campaign to Identify EBNA-1 Inhibitors project).Confirmatorydepositor-specified cross reference
2361Fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of the Epstein-Barr virus-encoded protein, ZTAScreeningsame project related to Summary assay
2362Counterscreen for inhibitors of ZTA: fluorescence polarization-based biochemical high throughput assay to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1)Screeningsame project related to Summary assay
488998Late stage counterscreen results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): fluorescence polarization-based biochemical dose response assay for inhibitors of the Epstein-Barr virus-encoded protein, ZTAConfirmatorysame project related to Summary assay
489000Late stage results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Fluorescence polarization-based biochemical dose response assay for EBNA-1 inhibitorsConfirmatorysame project related to Summary assay
493077Late stage assay provider counterscreen results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Fluorescence polarization-based biochemical dose response assay for inhibitors of the Epstein-Barr virus-encoded protein, ZTAConfirmatorysame project related to Summary assay
588762Late stage assay provider results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Cell-based luciferase-based dose response assay for EBNA-1 inhibitorsConfirmatorysame project related to Summary assay
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Paul Lieberman, Wistar Institute
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: I R21 NS063906-01
Grant Proposal PI: Paul Lieberman, Wistar Institute
External Assay ID: EBNA1_INH_FP_IC50 MDRUN Assay Provider

Name: Late stage assay provider results for the probe development effort to identify inhibitors of the Epstein-Barr virus nuclear antigen 1 (EBNA-1): Fluorescence polarization-based biochemical dose response assay for EBNA-1 inhibitors.

Description:

During each cell cycle in eukaryotes, the genome must be completely replicated and this replication must begin at the correct time and site (initiation site or origin) (1). Pathogenic viruses often take advantage of this cellular precision to maintain replication of their own genome. The Epstein-Barr virus (EBV) is an orally-transmitted herpes virus associated with numerous human neoplasms (2) that infects over 90% of the world's population (3). Latent EBV infection stimulates B cell proliferation in vitro, and can lead to B cell transformation (4). Following infection, the EBV genome is maintained in the host cell as a plasmid that is replicated by machinery comprised of several host proteins and a sole EBV protein, the EBV nuclear antigen 1 (EBNA-1). This EBNA-1 protein is required for replication of the EBV DNA genome. EBNA-1 is a DNA-binding protein that binds to an EBV sequence called the viral origin of replication plasmid, OriP (5). Studies demonstrating a role for EBV in Burkitt's lymphoma and Hodgkin's disease (6), multiple sclerosis (7), lupus (8), infectious mononucleosis (9), and nasopharyngeal carcinoma (10), combined with the recent discovery that EBNA-1 induces DNA double strand breaks (11), suggest that inhibitors of EBNA-1 may serve useful for understanding virus-cell interactions, virus-mediated cellular transformation, and as therapies for EBV-associated pathologies.

References:

1. Norseen, J, Thomae, A, Sridharan, V, Aiyar, A, Schepers, A and Lieberman, PM, RNA-dependent recruitment of the origin recognition complex. EMBO J, 2008. 27(22): p. 3024-35.
2. Carbone, A, Gloghini, A and Dotti, G, EBV-associated lymphoproliferative disorders: classification and treatment. Oncologist, 2008. 13(5): p. 577-85.
3. Schulz, TF and Cordes, S, Is the Epstein-Barr virus EBNA-1 protein an oncogene? Proc Natl Acad Sci U S A, 2009. 106(7): p. 2091-2.
4. Thorley-Lawson, DA and Babcock, GJ, A model for persistent infection with Epstein-Barr virus: the stealth virus of human B cells. Life Sci, 1999. 65(14): p. 1433-53.
5. Kirchmaier, AL and Sugden, B, Dominant-negative inhibitors of EBNA-1 of Epstein-Barr virus. J Virol, 1997. 71(3): p. 1766-75.
6. Hammerschmidt, W and Sugden, B, Epstein-Barr virus sustains Burkitt's lymphomas and Hodgkin's disease. Trends Mol Med, 2004. 10(7): p. 331-6.
7. Lunemann, JD, Kamradt, T, Martin, R and Munz, C, Epstein-barr virus: environmental trigger of multiple sclerosis? J Virol, 2007. 81(13): p. 6777-84.
8. Harley, JB, Harley, IT, Guthridge, JM and James, JA, The curiously suspicious: a role for Epstein-Barr virus in lupus. Lupus, 2006. 15(11): p. 768-77.
9. Vetsika, EK and Callan, M, Infectious mononucleosis and Epstein-Barr virus. Expert Rev Mol Med, 2004. 6(23): p. 1-16.
10. Raab-Traub, N, Epstein-Barr virus in the pathogenesis of NPC. Semin Cancer Biol, 2002. 12(6): p. 431-41.
11. Gruhne, B, Sompallae, R, Marescotti, D, Kamranvar, SA, Gastaldello, S and Masucci, MG, The Epstein-Barr virus nuclear antigen-1 promotes genomic instability via induction of reactive oxygen species. Proc Natl Acad Sci U S A, 2009. 106(7): p. 2313-8.

Keywords:

late stage, SAR, purchase, synthesis, assay provider, low throughput, EBNA-1, EBNA1, EBNA, Epstein-Barr virus, EBV, herpes virus, DNA virus, human herpes virus 4, HHV4, lymphoma, cancers, inhibitors, inhibition, antagonists, fluorescence polarization, FP, fluorescence, biochemical, dose response, Kansas, KUSCC, Kansas University Specialized Chemistry Center, Scripps, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Protocol
Assay Overview:
The purpose of this assay is to determine dose response curves for powder samples of compounds identified as possible EBNA-1 inhibitor probe candidates. This assay assesses the ability of compounds to inhibit the binding of EBNA-1 protein to DNA. In this biochemical assay, recombinant EBNA-1 DNA binding domain (DBD) (aa 448-610) purified from E. coli is incubated with a Cyanine (Cy5)-labeled 36-bp hairpin DNA oligonucleotide probe in the presence of test compounds. The reaction is excited with linear polarized light and the intensity of the emitted light is measured as the polarization value. As designed, compounds that act as EBNA inhibitors will prevent EBNA-DNA binding, thereby increasing the proportion of free DNA in the well, resulting in low fluorescence polarization in the well. This assay was performed by the assay provider.
Protocol Summary:
Please contact the assay provider for assay details.
PubChem Activity Outcome and Score:
Compounds with an IC50 greater than 10 uM were considered inactive. Compounds with an IC50 equal to or less than 10 uM were considered active.
Activity score was then ranked by the potency, with the most potent compounds assigned the highest activity scores.
The PubChem Activity Score range for active compounds is 100-100, and for inactive compounds 89-0.
List of Reagents:
Please contact the assay provider for assay details.
Comment
This assay was performed by the assay provider. Replicate values and raw data were not provided. This assay may have been run as two or more separate campaigns, each campaign testing a unique set of compounds. Possible artifacts of this assay can include, but are not limited to: dust or lint and compounds that modulate fluorescence. All test compound concentrations reported above and below are nominal; the specific test concentration(s) for a particular compound may vary based upon the actual sample provided by the Kansas University Specialized Chemistry Center.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Biochemical
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Qualifier [IC50]Activity Qualifier identifies if the resultant data IC50 came from a fitted curve or was determined manually to be less than or greater than its listed IC50 concentration.String
2IC50*The concentration at which 50 percent of the activity in the inhibitor assay is observed; (IC50) shown in uM.FloatμM
3Hill SlopeThe variable HillSlope describes the steepness of the curve. This variable is called the Hill slope, the slope factor, or the Hill coefficient. If it is positive, the curve increases as X increases. If it is negative, the curve decreases as X increases. A standard sigmoid dose-response curve (previous equation) has a Hill Slope of 1.0. When HillSlope is less than 1.0, the curve is more shallow. When HillSlope is greater than 1.0, the curve is steeper. The Hill slope has no units.Float
4EBNA1 % ResponseValue of % response.Float%
5EBNA1 % Response Range (+/-)Range identifies the value range for the EBNA % response.Integer

* Activity Concentration.
Additional Information
Grant Number: I R21 NS063906-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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