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BioAssay: AID 449731

Summary of the probe development effort to identify inhibitors of AddAB recombination protein complex

Name: Summary of the probe development effort to identify inhibitors of AddAB recombination protein complex. ..more
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AID: 449731
Data Source: The Scripps Research Institute Molecular Screening Center (ADDAB_INH_SUMMARY)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-07-07
Modify Date: 2012-12-17
Targets
Related Experiments
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AIDNameTypeComment
435030Absorbance-based primary bacterial cell-based high throughput screening assay to identify inhibitors of AddAB recombination protein complexScreeningdepositor-specified cross reference: Primary screen (AddAB inhibitors in singlicate)
449728Counterscreen for inhibitors of AddAB: absorbance-based bacterial cell-based high throughput screening assay to identify inhibitors of bacterial viabilityScreeningdepositor-specified cross reference: Counterscreen (Cytotoxicity in singlicate)
488942Absorbance-based bacterial cell-based high throughput confirmation assay for inhibitors of AddAB recombination protein complexScreeningdepositor-specified cross reference: Confirmation screen (AddAB inhibitors in triplicate)
488955Counterscreen for AddAB inhibitors: absorbance-based high throughput cell-based assay to identify inhibitors of RecBCDScreeningdepositor-specified cross reference: Counterscreen (RecBCD inhibitors in triplicate)
488956Counterscreen for AddAB inhibitors: absorbance-based bacterial cell-based high throughput confirmation assay for inhibitors of bacterial viabilityScreeningdepositor-specified cross reference: Counterscreen (bacterial viability inhibitors in triplicate)
492957Counterscreen for AddAB inhibitors: absorbance-based bacterial cell-based high throughput dose response assay to identify inhibitors of RecBCDConfirmatorydepositor-specified cross reference: Dose response counterscreen (RecBCD inhibitors in triplicate)
492958Counterscreen for AddAB inhibitors: absorbance-based bacterial cell-based high throughput dose response assay for inhibitors of bacterial viabilityConfirmatorydepositor-specified cross reference: Dose response counterscreen (bacterial viability inhibitors in triplicate)
492959Absorbance-based bacterial cell-based high throughput dose response assay for inhibitors of AddAB recombination protein complexConfirmatorydepositor-specified cross reference: Dose response (AddAB inhibitors in triplicate)
504677Late stage results for the probe development effort to identify inhibitors of AddAB recombination protein complex: Absorbance-based bacterial cell-based dose response assay for inhibitors of AddAB recombination protein complexConfirmatorydepositor-specified cross reference: Late stage dose response (AddAB recombination protein complex inhibitors in triplicate)
504678Late stage counterscreen results for the probe development effort to identify inhibitors of AddAB recombination protein complexes: absorbance-based bacterial cell-based dose response assay for inhibitors of bacterial viabilityConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (bacterial viability inhibitors in triplicate)
504679Late stage counterscreen results for the probe development effort to identify inhibitors of AddAB recombination protein complex: absorbance-based bacterial cell-based dose response assay to identify inhibitors of RecBCDConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (RecBCD inhibitors in triplicate)
602421Late stage assay provider assay for AddAB inhibitors: Radioactivity-based biochemical dose response assay to identify inhibitors of the nuclease activity of purified AddABConfirmatorydepositor-specified cross reference: Late stage dose response (purified AddAB nuclease activity inhibitors)
602422Late stage assay provider assay for AddAB inhibitors: Radioactivity-based biochemical assay to identify inhibitors of the nuclease activity of purified AddAB (100 micromolar dose)Otherdepositor-specified cross reference: Late stage assay (purified AddAB nuclease activity inhibitors)
623916Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony-formation assay to identify inhibitors of the viability of V66 E. coli in Hfr recombination assaysOtherdepositor-specified cross reference: Late stage counterscreen (viability of V66 E. coli in Hfr recombination assays inhibitors)
623917Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony-formation dose response assay to identify inhibitors of the viability of V66 E. coli in Hfr recombination assaysOtherdepositor-specified cross reference: Late stage dose response counterscreen (viability of V66 E. coli in Hfr recombination assays inhibit
623918Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony formation assay to identify inhibitors of the recombination-promoting activity of RecBCD in V66 E. coli (dose response)Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (recombination-promoting activity of RecBCD in V66 E. coli in
623919Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony formation assay to identify inhibitors of the recombination-promoting activity of RecBCD in V66 E. coli (100uM)Otherdepositor-specified cross reference: Late stage counterscreen (recombination-promoting activity of RecBCD in V66 E. coli inhibitors at 10
623920Late stage assay provider counterscreen for AddAB inhibitors: Radioactivity-based biochemical dose response assay to identify inhibitors of the nuclease activity of purified RecBCD enzymeConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (nuclease activity of purified RecBCD enzyme inhibitors)
623921Late stage assay provider counterscreen for AddAB inhibitors: Radioactivity-based biochemical assay to identify inhibitors of the nuclease activity of purified RecBCD enzyme (at 100 micromolar)Otherdepositor-specified cross reference: Late stage counterscreen (nuclease activity of purified RecBCD enzyme inhibitors at 100 uM)
623922Late stage assay provider counterscreen for AddAB inhibitors: Radioactivity-based biochemical assay to identify inhibitors of the nuclease activity of purified RecBCD enzyme (at 50 micromolar)Otherdepositor-specified cross reference: Late stage counterscreen (nuclease activity of purified RecBCD enzyme inhibitors at 50 uM)
623937Late stage assay provider counterscreen for AddAB inhibitors: Radioactivity-based biochemical assay to identify inhibitors of the helicase/Chi cutting activity of purified RecBCDOtherdepositor-specified cross reference: Late stage counterscreen (helicase/Chi cutting activity of purified RecBCD inhibitors)
623942Late stage assay provider counterscreen for AddAB inhibitors: Radioactivity-based biochemical dose response assay to identify inhibitors of the helicase/Chi cutting activity of purified RecBCDConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (helicase/Chi cutting activity of purified RecBCD inhibitors)
623954Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony-formation assay to identify inhibitors of recombination of E. coli transformed with p(addAB) and p(recA) (100 micromolar compound dose)Otherdepositor-specified cross reference: Late stage counterscreen (recombination of E. coli transformed with p(addAB) and p(recA) inhibitors)
623956Late stage assay provider Counterscreen for AddAB inhibitors: Cell-based colony-formation assay to identify inhibitors of recombination of E. coli transformed with p(addAB) and p(recA) (20 micromolar compound dose)Otherdepositor-specified cross reference: Late stage counterscreen (recombination of E. coli transformed with p(addAB) and p(recA) inhibitors)
651942Late stage results for the probe development effort to identify inhibitors of AddAB recombination protein complex: Absorbance-based bacterial cell-based dose response assay for inhibitors of AddAB recombination protein complex (ROUND 2)Confirmatorydepositor-specified cross reference: Late stage dose response assay (AddAB recombination protein complex inhibitors in triplicate)
651943Late stage counterscreen results for the probe development effort to identify inhibitors of AddAB recombination protein complexes: absorbance-based bacterial cell-based dose response assay for inhibitors of bacterial viability (ROUND 2)Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (bacterial viability inhibitors in triplicate)
651944Late stage counterscreen results for the probe development effort to identify inhibitors of AddAB recombination protein complex: absorbance-based bacterial cell-based dose response assay to identify inhibitors of RecBCD (ROUND 2)Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (RecBCD inhibitors in triplicate)
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Gerald R. Smith, Fred Hutchinson Cancer Research Center
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: GM031693
Grant Proposal PI: Gerald R. Smith
External Assay ID: ADDAB_INH_SUMMARY

Name: Summary of the probe development effort to identify inhibitors of AddAB recombination protein complex.

Description:

Helicobacter pylori infects approximately half of the world's population and is responsible for inducing chronic gastric inflammation that can progress to gastric cancer (1). At the cellular level, Helicobacter pylori infection of the human stomach is associated with inflammation that elicits DNA damage in both bacterial and host cells (2). This DNA damage must be repaired in order for the bacteria to persist. The H. pylori AddAB helicase-exonuclease is required for DNA repair and efficient stomach colonization (3), and inhibitors of this enzyme may be useful antibacterial drugs for treating these infections. The AddAB class of enzymes is closely related to the RecBCD class of helicase-nucleases; both classes are widely distributed in bacteria but appear to be absent in eukaryotes (4). The protein complex functions in DNA repair by directing free DNA ends into the homologous recombination pathway (5). As a result, the identification of inhibitors of AddAB may be useful tools for elucidating the role of AddAB and RecBCD in bacterial recombination and as potential novel antibiotics with few off-target effects.

Summary of Probe Development Effort:

This probe development effort is focused on the identification of ADDAB inhibitors. All AIDs that contain results associated with this project can be found in the "Related Bioassays" section of this Summary AID.

References:

1. Fox JG, Wang TC. Inflammation, atrophy, and gastric cancer. J Clin Invest. 2007 Jan;117(1):60-9.
2. Ernst P. Aliment Pharmacol Ther. 1999 Mar;13 Suppl 1:13-8. Review article: the role of inflammation in the pathogenesis of gastric cancer.
3. Dillingham MS, Kowalczykowski SC. RecBCD enzyme and the repair of double-stranded DNA breaks. Microbiol Mol Biol Rev. 2008 Dec;72(4):642-71.
4. Amundsen SK, Fero J, Hansen LM, Cromie GA, Solnick JV, Smith GR, Salama NR, Helicobacter pylori AddAB helicase-nuclease and RecA promote recombination-related DNA repair and survival during stomach colonization. Mol Microbiol, 2008. 69(4): p. 994-1007.
5. Chedin F. and Kowalczykowski S.C. A novel family of regulated helicases/nucleases from Gram-positive bacteria: insights into the initiation of DNA recombination, Mol. Microbiol. 43 (2002), pp. 823-834.

Keywords:

Summary, Summary AID, helicase, nuclease, exonuclease, ATP-dependent nuclease, AddAB, ADDAB, AddAB complex, RecBCD enzyme, beta subunit, gamma chain, alpha chain, Escherichia coli, E. coli, bacteria, Helicobacter pylori, phage, T4, DNA, dsDNA, DNA damage, DNA repair, DNA binding, ATP-binding, homologous recombination, recombination, Chi, inhibition, inhibitor, optical density, OD, absorbance, HTS, high throughput screen, primary, 1536, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML328
PubChem Substance ID (SID) for probe 1: 144186606
PubChem Compound ID (CID) for probe 1: 1517823
Probe type for probe 1: Inhibitor
IC50/EC50 (nM) for probe 1: 1000
Target for probe 1: AddAB; RecBCD (gi: 15645703,15646160)
Disease relevance for probe 1: Bacterial Infection
Anti-target for probe 1: E. Coli V66 Viability
Fold selectivity for probe 1: 34
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK148492/ (ID: 3040622)
Grant number for probe 1: GM031693
PubMed Publication ID (PMID) for probe 1: 22443934
NCBI Book chapter title for probe 1: ML328: A Novel Dual Inhibitor of Bacterial AddAB and RecBCD Helicase-nuclease DNA Repair Enzymes
Additional Information
Grant Number: GM031693

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