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BioAssay: AID 492969

Summary of the probe development efforts to identify inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2)

Name: Summary of the probe development efforts to identify inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2). ..more
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 Tested Compounds
 Tested Compounds
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Probe(1)
 
 
Active(1)
 
 
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 Tested Substances
All(1)
 
 
Probe(1)
 
 
Active(1)
 
 
AID: 492969
Data Source: The Scripps Research Institute Molecular Screening Center (PAFAH2_INH_SUMMARY)
BioAssay Type: Panel, Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-12-08
Modify Date: 2011-10-18

Data Table ( Complete ):           View Data Probes    View Active Data    View All Data
Target
BioActive Compound: Chemical Probe: 1    Active: 1
Related Experiments
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AIDNameTypeComment
463082Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH)Screeningdepositor-specified cross reference: Counterscreen (pPAFAH inhibitors in singlicate)
492956Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2)Screeningdepositor-specified cross reference: Primary screen (PAFAH2 inhibitors in singlicate)
493030Fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2)Screeningdepositor-specified cross reference: Confirmation screen (PAFAH2 inhibitors in triplicate)
504483Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) counterscreen assay to assess selectivity against anti-target pPAFAH in vitroOtherdepositor-specified cross reference: Late stage counterscreen (ABPP selectivity against anti-target pPAFAH)
504486Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: LC-MS/MS assay to assess binding of compounds to active siteOtherdepositor-specified cross reference: Late stage assay (LC-MS/MS, assess binding of compounds to active site)
504491Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) inhibition and selectivity to assess carbamate vs. triazole urea scaffoldOtherdepositor-specified cross reference: Late stage assay (ABPP inhibition and selectivity, carbamate vs. triazole urea scaffold)
504494Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) IC50Confirmatorydepositor-specified cross reference: Late stage dose response (PAFAH2 inhibitors in triplicate)
504495Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: fluorescence-based cell-based gel-based Activity-Based Protein Profiling (ABPP) inhibitionOtherdepositor-specified cross reference: Late stage assay (ABPP inhibition in singlicate)
504496Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: fluorescence-based cell-based gel-based Activity-Based Protein Profiling (ABPP) IC50Confirmatorydepositor-specified cross reference: Late stage dose response (PAFAH2 inhibitors in triplicate)
504511Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compoundsConfirmatorydepositor-specified cross reference: Late stage dose response (cytotoxicity in triplicate)
504513Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) inhibition and selectivityOtherdepositor-specified cross reference: Late stage assay (ABPP inhibition and selectivity)
504519Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: LC-MS-based cell-based SILAC Activity-Based Protein Profiling (ABPP) for PAFAH2Otherdepositor-specified cross reference: Late stage assay (LC-MS SILAC ABPP for PAFAH2)
504527Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) general inhibition and selectivity of serine hydrolasesOtherdepositor-specified cross reference: Late stage assay (ABPP general inhibition and selectivity of serine hydrolases)
504531Late stage assay provider results from the probe development effort to identify inhibitors of PAFAH2: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) assay to assess selectivity against anti-target pPAFAH in situOtherdepositor-specified cross reference: Late stage assay (ABPP to assess selectivity against anti-target pPAFAH in situ)
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Brian J. Bahnson, University of Delaware
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: HL084366
Grant Proposal PI: Brian J. Bahnson, University of Delaware
External Assay ID: PAFAH2_INH_SUMMARY

Name: Summary of the probe development efforts to identify inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2).

Description:

Atherosclerosis is a process in which plaques, deposits of low density lipoproteins (LDL), lipid-laden macrophages and other inflammatory cells, calcium, and cellular debris build up in the inner lining of an artery. These plaques interfere with blood flow, damage the arterial wall, and eventually rupture, causing debris to migrate downstream, leading to myocardial infarction or stroke (1-4). Human platelet activating factor acetylhydrolase (pPAFAH) is a Ca2+ independent phospholipase A2 (PLA2) identified in human plasma as the enzyme responsible for the hydrolysis/inactivation of platelet activating factor (PAF) (5, 6), a potent pro-inflammatory phospholipid signaling molecule (7). PAFAH catalyses the hydrolysis of the acetyl group at the sn-2 position of the glycerol backbone of PAF converting it to lyso-PAF (8-9). PAFAH has a catalytic cysteine residue and is consequently sensitive to broadly reactive thiol alkylating agents, including N-ethylmaleimide (10); however, selective inhibitors for PAFAH have not yet been identified. Inhibiting PAFAH may thus offer a new therapeutic strategy for cancer. Development of a selective inhibitor would also aid in the investigation into PAFAH involvement in the dysregulated biochemical pathways that support tumorigenesis. As a result, the identification of inhibitors of PAFAH would help to elucidate the physiological role of this enzyme and its contribution to atherosclerosis, cancer, and other inflammatory pathologies (11).

Summary of Probe Development Effort:

This probe development effort is focused on the identification of inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2). All AIDs that contain results associated with this project can be found in the "Related Bioassays" section of this Summary AID.

References:

1. Karasawa, K., Harada, A., Satoh, N., Inoue, K., and Setaka, M. (2003) Plasma platelet activating factor acetylhydrolase (PAF-AH), Prog Lipid Res 42, 93-114.
2. Leitinger, N. (2005) Oxidized phospholipids as triggers of inflammation in atherosclerosis, Molecular Nutrition & Food Research 49, 1063-1071.
3. Blank, M. L., Lee, T., Fitzgerald, V., and Snyder, F. (1981) A specific acetylhydrolase for 1-alkyl-2- acetyl-sn glycero-3-phosphocholine (a hypotensive and platelet-activating lipid), J Biol Chem 256, 175-178.
4. Farr, R. S., Cox, C. P., Wardlow, M. L., and Jorgensen, R. (1980) Preliminary studies of an acid labile factor (ALF) in human sera that inactivates platelet-activating factor (PAF), Clin Immunol Immunopathol 15, 318-330.
5. Zimmerman, G. A., McIntyre, T. M., Prescott, S. M., and Stafforini, D. M. (2002) The plateletactivating factor signaling system and its regulators in syndromes of inflammation and thrombosis, Crit Care Med 30, S294-301.
6. Anderson, J. L. (2008) Lipoprotein-associated phospholipase A2: an independent predictor of coronary artery disease events in primary and secondary prevention, Am J Cardiol 101, 23F-33F.
7. Sudhir, K. (2005) Clinical review: Lipoprotein-associated phospholipase A2, a novel inflammatory biomarker and independent risk predictor for cardiovascular disease, J Clin Endocrinol Metab 90, 3100-3105.
8. Wilensky, R. L., and Macphee, C. H. (2009) Lipoprotein-associated phospholipase A(2) and atherosclerosis, Curr Opin Lipidol 20, 415-420.
9. Karabina, S. A., and Ninio, E. (2006) Plasma PAF-acetylhydrolase: an unfulfilled promise?, Biochim Biophys Acta 1761, 1351-1358.
10. Samanta, U., and Bahnson, B. J. (2008) Crystal structure of human plasma platelet-activating factor acetylhydrolase: structural implication to lipoprotein binding and catalysis, J Biol Chem 283, 31617-31624.
11. Blackie, J. A., Bloomer, J. C., Brown, M. J. B., Cheng, H. Y., Hammond, B., Hickey, D. M. B., Ife, R. J., Leach, C. A., Lewis, V. A., Macphee, C. H., Milliner, K. J., Moores, K. E., Pinto, I. L., Smith, S. A., Stansfield, I. G., Stanway, S. J., Taylor, M. A., and Theobald, C. J. (2003) The identification of clinical candidate SB-480848: A potent inhibitor of lipoprotein-associated phospholipase A(2), Bioorganic & Medicinal Chemistry Letters 13, 1067-1070.

Keywords:

Summary, Summary AID, FLJ26025; HSD-PLA2, PAFAH2, 40kDa, pPAFAH, PAFAH1b2, PAFAH1b3, hydrolase, lipase, platelet activating factor acetylhydrolase 2, FP-Rh, atherosclerosis, heart disease, fluorescence polarization, FP, fluorescence, inhibit, inhibition, inhibitor, primary, primary screen, HTS, 1536, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Panel Information
AIDs
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1AID 504494platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] [gi:225579137]
Taxonomy id: 10090
Gene id: 100163
2AID 504511_1cytotoxicity, serum-free media
3AID 504511_2cytotoxicity, media plus FCS

§ Panel component ID.
Result Definitions
TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1ML #Unique alphanumeric identifier assigned to a chemical probe molecule within the Molecular Libraries Probe Production Centers Network (MLPCN).String
2IC50 [AID 504494] *Data from AID 504494. The value for concentration at which 50% inhibition is observed; IC50 shown in uM.1platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus]FloatμM
3CC50 [AID 504511_1] *Data from AID 504511, panel 1. The value for concentration at which 50% of surviving cells are observed; CC50 shown in micromolar.2FloatμM
4Qualifier CC50 [AID 504511_2] Data from AID 504511, panel 2. Activity Qualifier identifies if the resultant data CC50 came from a fitted curve or was determined manually to be less than or greater than its listed CC50 concentration.2String
5CC50 [AID 504511_2] *Data from AID 504511, panel 2. The value for concentration at which 50% of surviving cells are observed; CC50 shown in micromolar.3FloatμM

* Activity Concentration. § Panel component ID.
Additional Information
Grant Number: HL084366

Classification
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