Name: Summary of probe development efforts to identify inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH). ..more
Target
Depositor Specified Assays
Show more |
| AID | Name | Type | Comment |
|---|---|---|---|
| 463082 | Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH) | screening | Primary screen (pPAFAH inhibitors in singlicate) |
| 463230 | Fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH; PLA2G7) | screening | Confirmation screen (pPAFAH inhibitors in triplicate) |
| 492956 | Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of human platelet activating factor acetylhydrolase 2 (PAFAH2) | screening | Counterscreen (PAFAH2 inhibitors in singlicate) |
| 588474 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of pPAFAH | other | Late stage assay (ABPP inhibition of pPAFAH in singlicate) |
| 588766 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based dose response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of human pPAFAH | confirmatory | Late stage dose response (pPAFAH inhibitors in triplicate) |
| 588767 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of PAFAH2 | other | Late stage counterscreen (PAFAH2 inhibitors) |
| 588768 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compounds | confirmatory | Late stage dose response (Cytotoxicity in 5 replicates) |
| 588769 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based dose response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) assay for HTS compounds | confirmatory | Late stage dose response (ABPP for HTS compounds in triplicate) |
| 588770 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) assay for SAR compounds | confirmatory | Late stage assay (ABPP for SAR compounds in triplicate) |
| 588773 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition and selectivity of HTS compounds in a complex proteome | other | Late stage assay (ABPP inhibition and selectivity of HTS compounds in a complex proteome) |
| 588774 | Late stage assay provider results from the probe development effort to identify inhibitors of plasma platelet activating factor acetylhydrolase (pPAFAH): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition and selectivity of SAR compounds in a complex proteome | other | Late stage assay (ABPP inhibition and selectivity of SAR compounds in a complex proteome) |
| 588785 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: LC-MS/MS-based biochemical ABPP-MudPIT assay to determine selectivity of test compounds in the mouse brain membrane proteome | other | Late stage results (LC-MS/MS-based biochemical ABPP-MudPIT selectivity in the mouse brain membrane proteome) |
| 588787 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: LC-MS/MS-based biochemical ABPP-MudPIT assay to determine selectivity of test compounds in the mouse brain soluble proteome | other | Late stage results (LC-MS/MS-based biochemical ABPP-MudPIT selectivity in the mouse brain soluble proteome) |
| 588788 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: LC-MS/MS-based biochemical assay to determine binding mode of test compounds | other | Late stage results (LC-MS/MS-based biochemical, binding mode) |
| 588789 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of pPAFAH in vivo | other | Late stage results (ABPP inhibition of pPAFAH in vivo) |
| 588817 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of pPAFAH in situ | other | Late stage assay (ABPP inhibition of pPAFAH in situ) |
| 588823 | Late stage assay provider results from the probe development effort to identify inhibitors of pPAFAH: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of pPAFAH and selectivity analysis in vivo | other | Late stage assay (ABPP inhibition of pPAFAH and selectivity analysis in vivo) |
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: PPAFAH_INH_SUMMARY
Name: Summary of probe development efforts to identify inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH).
Description:
Atherosclerosis is a process in which plaques, deposits of low density lipoproteins (LDL), lipidladen 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 proinflammatory 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 the plasma platelet activating factor acetylhydrolase (pPAFAH). 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-snglycero-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 platelet-activating 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, PLA2G7, PAFAH, pPAFAH, PPAFAH, LP-LPA2, PAFAH2, PAFAH1b2, PAFAH1b3, hydrolase, plasma platelet activating factor acetylhydrolase, FP-Rh, atherosclerosis, heart disease, cancer, fluorescence polarization, FP, antagonist, inhibitor, inhibit, primary, HTS, high throughput screening, 1536, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
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: PPAFAH_INH_SUMMARY
Name: Summary of probe development efforts to identify inhibitors of the plasma platelet activating factor acetylhydrolase (pPAFAH).
Description:
Atherosclerosis is a process in which plaques, deposits of low density lipoproteins (LDL), lipidladen 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 proinflammatory 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 the plasma platelet activating factor acetylhydrolase (pPAFAH). 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-snglycero-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 platelet-activating 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, PLA2G7, PAFAH, pPAFAH, PPAFAH, LP-LPA2, PAFAH2, PAFAH1b2, PAFAH1b3, hydrolase, plasma platelet activating factor acetylhydrolase, FP-Rh, atherosclerosis, heart disease, cancer, fluorescence polarization, FP, antagonist, inhibitor, inhibit, primary, HTS, high throughput screening, 1536, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Panel Information
AIDs
§ Panel component ID.
| Data Table(Active) Data Table(All) | Show more |
| PID§ | Name | Substance | Panel Targets | Description | Additional Information | ||
|---|---|---|---|---|---|---|---|
| Active | Inactive | ||||||
| 1 | AID 588766 | platelet-activating factor acetylhydrolase precursor [Homo sapiens] [gi:270133071] | Taxonomy id: 9606 Gene id: 7941 | ||||
| 2 | AID 588770 | Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [Mus musculus] [gi:14715116] | Taxonomy id: 10090 Gene id: 27226 | ||||
| 3 | AID 588768_1 | ||||||
| 4 | AID 588768_2 | ||||||
§ Panel component ID.
Result Definitions
| TID | Name | Description | PID§ | Panel Targets | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||||
| Score | The BioAssay activity ranking score |  | Integer | |||||
| 1 | ML # | Unique alphanumeric identifier assigned to a chemical probe molecule within the Molecular Libraries Probe Production Centers Network (MLPCN). | String | |||||
| 2 | Average IC50 [AID 588766] | The average concentration at which 50 percent of the activity in the antagonist assay is observed; (IC50) shown in micromolar. | 1 | platelet-activating factor acetylhydrolase precursor [Homo sapiens] | | Float | μM | |
| 3 | Average IC50 [AID 588770] | The average concentration at which 50 percent of the activity in the antagonist assay is observed; (IC50) shown in micromolar. | 2 | Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [Mus musculus] | | Float | μM | |
| 4 | CC50 [AID 588768_1] | The value for the concentration at which 50% of surviving cells are observed in serum-free medium; CC50 shown in micromolar. | 3 | | Float | μM | ||
| 5 | CC50 [AID 588768_2] | The value for the concentration at which 50% of surviving cells are observed in serum-containing medium; CC50 shown in micromolar. | 4 | | Float | μM |
Additional Information
Grant Number: HL084366
Classification
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