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BioAssay: AID 2202

Summary of probe development efforts to identify inhibitors of lysophospholipase 1 (LYPLA1).

Reversible protein phosphorylation networks play essential roles in most cellular processes. While over 500 kinases catalyze protein phosphorylation, only two enzymes, PP1 and PP2a, are responsible for >90% of all serine/ threonine phosphatase activity (1). Phosphatases, unlike kinases, achieve substrate specificity through complex subunit assembly and post-translational modifications rather more ..
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 Tested Compounds
 Tested Compounds
All(2)
 
 
Probe(2)
 
 
Active(2)
 
 
 Tested Substances
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Probe(2)
 
 
Active(2)
 
 
AID: 2202
Data Source: The Scripps Research Institute Molecular Screening Center (LYPLA1_INH_POST-PRUN_SUMMARY)
BioAssay Type: Panel, Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-12-12
Modify Date: 2013-02-07

Data Table ( Complete ):           Probe    Active    All
Target
BioActive Compounds: Chemical Probe: 2    Active: 2
Depositor Specified Assays
Show more
AIDNameTypeProbeComment
2174Counterscreen for PME1 inhibitors: fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of lysophospholipase 1 (LYPLA1).screening Screening assay (LYPLA1 inhibitors).
493105Assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition of recombinant and endogenous enzymeother Late stage screening assay (LYPLA1 and LYPLA2 dual inhibitors)
493108Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: fluorescence-based cell-based inhibitionother Late stage screening assay (LYPLA1 and LYPLA2 dual inhibitors)
493109Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: LC-MS/MS assay to assess binding of compounds to active siteother Late stage assay (LC-MS/MS binding of compounds to LYPLA1 active site)
493110Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: Gel-based Activity-Based Protein Profiling (ABPP) IC50 for LYPLA1 and LYPLA2confirmatory Late stage dose response (LYPLA1 and LYPLA2 dual inhibitors)
493111Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition and selectivityother Late stage counterscreen assay (LYPLA1, LYPLA2, ABHD11, ESD, AND APEH inhibitors)
493154Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1 and LYPLA2: Gel-based Activity-Based Protein Profiling (ABPP) IC50 for off-target ABHD11confirmatory Late stage dose response counterscreen (ABHD11 inhibitors)
493161Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compoundsconfirmatory Late stage dose response counterscreen (Cytotoxicity in quadruplicate)
504482Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: fluorescence-based cell-based gel-based Activity-Based Protein Profiling (ABPP) IC50 for anti-target ABHD11confirmatory Late stage dose response counterscreen (ABHD11 inhibitors in triplicate)
504498Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: LC-MS/MS assay to assess binding of compounds to active site of anti-target ABHD11other Late stage assay (LC-MS/MS assess binding of compounds to active site of anti-target ABHD11)
504505Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: fluorescence-based cell-based gel-based Activity-Based Protein Profiling (ABPP) percent inhibition for anti-target ABHD11other Late stage assay (ABPP inhibition for anti-target ABHD11)
504507Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) IC50 for anti-target ABHD11 Set 2confirmatory Late stage dose response counterscreen (ABHD11 inhibitors in triplicate)
504510Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compounds set 2confirmatory Late stage dose response counterscreen (cytotoxicity in triplicate)
504520Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) inhibition and selectivityother Late stage assay (ABPP inhibition and selectivity)
504522Late stage assay provider results from the probe development effort to identify inhibitors of LYPLA1: LC-MS-based cell-based SILAC Activity-Based Protein Profiling (ABPP) for anti-target ABHD11other Late stage (LC-MS-based cell-based SILAC ABPP for anti-target ABHD11)
504892Late stage assay provider results from the probe development effort to identify inhibitors of ABHD11: Fluorescence-based biochemical gel-based Activity-Based Protein Profiling (ABPP) inhibition of the human isoform of ABHD11other Late stage assay (human isoform of ABHD11 inhibitors)
651978Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1: LCMS-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis in vitroother Late stage assay (SILAC selectivity analysis in vitro)
651979Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1: LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis in situother Late stage assay (SILAC selectivity analysis in situ)
651985Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP evaluation of activity in vivoother Late stage assay (LYPLA1 and LYPLA2 inhibitors activity in vivo)
651986Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP evaluation of activity in situother Late stage assay (LYPLA1 and LYPLA2 inhibitors activity in situ)
651987Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP gel filtration assay to assess binding modeother Late stage assay (binding mode)
651988Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition and selectivityother Late stage assay (inhibition and selectivity)
651990Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Fluorescence-based biochemical dose-response gel-based ABPPconfirmatory Late stage dose-response (LYPLA1 and LYPLA2 inhibitors in triplicate)
651991Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1 and LYPLA2: Absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compoundsconfirmatory Late stage dose-response (cytotoxicity 6 replicates)
651998Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1: Substrate-based fluorescence-based biochemical determination of kinetic parametersconfirmatory Late stage assay (LYPLA1 kinetic parameters in quadruplicate)
652003Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1: fluorescence-based biochemical dose-response assayconfirmatory Late stage dose-response (LYPLA1 inhibitors in quadruplicate)
652018Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA1: LCMS-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis in vitro, Set 2other Late stage assay (selectivity analysis in vitro)
2233Counterscreen for PME1 inhibitors: fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of lysophospholipase 1 (LYPLA1).screening
651981Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: LCMS-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis in vitroother
2143Summary of probe development efforts to identify inhibitors of Protein Phosphatase Methylesterase 1 (PME-1).summary3
652001Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: Substrate-based fluorescence-based biochemical determination of kinetic parametersconfirmatory
652004Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: fluorescence-based biochemical dose-response assayconfirmatory
652030Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition and selectivityother
651980Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis in situother
652029Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibitionother
743117 On Hold
743118 On Hold
743119 On Hold
743127 On Hold
743132 On Hold
743133 On Hold
743134 On Hold
743137 On Hold
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Ben Cravatt, TSRI
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 CA132630 Fast Track
Grant Proposal PI: Ben Cravatt, TSRI
External Assay ID: LYPLA1_INH_POST-PRUN_SUMMARY

Name: Summary of probe development efforts to identify inhibitors of lysophospholipase 1 (LYPLA1).

Description:

Reversible protein phosphorylation networks play essential roles in most cellular processes. While over 500 kinases catalyze protein phosphorylation, only two enzymes, PP1 and PP2a, are responsible for >90% of all serine/ threonine phosphatase activity (1). Phosphatases, unlike kinases, achieve substrate specificity through complex subunit assembly and post-translational modifications rather than number. PP2a, for example, typically exists as heterotrimer with diverse subunits that may combinatorially make as many as 70 different holoenzyme assemblies (2). Mutations in several of these PP2a subunits have been identified in human cancers, suggesting that PP2a may act as a tumor suppressor (3). Adding further complexity, several residues of the catalytic subunit of PP2a can be reversibly phosphorylated, and the C-terminal leucine residue can be reversibly methylated (4,5). Protein phosphatase methylesterase 1 (PME-1) is specifically responsible for demethylation of the carboxyl terminus (6).

Methylesterification is thought to control the binding of different subunits to PP2a, but little is known about physiological significance of this post-translational modification in vivo (7). Recently, PME-1 has been identified as a protector of sustained ERK pathway activity in malignant gliomas (8). In order to further elucidate the role of PP2a methylation in vivo, our lab has generated mice that lack PME-1 (PME-1 (-/-) mice) by targeted gene disruption (9). Unfortunately, PME-1 deletion resulted in perinatal lethality, underscoring the importance of PME-1 but hindering our biological studies. Biochemical elucidation of PME-1 and other protein modifiers such as lysophospholipases (LYPLA) would thus greatly benefit from the development of potent and selective chemical inhibitors (10).

Summary of Probe Development Effort:

This probe development effort is focused on the identification of LYPLA1 inhibitors (AID 2174). All AIDs that contain results associated with this project can be found in the "Related Bioassays" section of this Summary AID.

References:

1. Oliver, C. J., Shenolikar, S. (1998). Physiologic importance of protein phosphatase inhibitors. Front. Biosci. 3, D961-972.
2. Janssens, V., Goris, J. (2001). Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417-439.
3. Janssens, V., Goris, J., Van Hoof, C. (2005). PP2A: the expected tumor suppressor. Curr. Opin. Genet. Dev. 15, 34-41.
4. Chen, J., Martin, B. L., Brautigan, D. L. (1992). Regulation of protein serine-threonine phosphatase type-2A by tyrosine phosphorylation. Science 257, 1261-1264.
5. Favre, B., Zolnierowicz, S., Turowski, P., Hemmings, B. A. (1994). The catalytic subunit of protein phosphatase 2A is carboxyl-methylated in vivo. J. Biol. Chem. 269, 16311-16317.
6. Lee, J., Chen, Y., Tolstykh, T., Stock, J. (1996). A specific protein carboxyl methylesterase that demethylates phosphoprotein phosphatase 2A in bovine brain. Proc. Natl. Acad. Sci. U. S. A. 93, 6043-6047.
7. Wu, J., Tolstykh, T., Lee, J., Boyd, K., Stock, J. B., Broach, J. R. (2000). Carboxyl methylation of the phosphoprotein phosphatase 2A catalytic subunit promotes its functional association with regulatory subunits in vivo. Embo J. 19, 5672-5681.
8. Puustinen, P., Junttila, M. R., Vanhatupa, S., Sablina, A. A., Hector, M. E., Teittinen, K., Raheem, O., Ketola, K., Lin, S., Kast, J., Haapasalo, H., Hahn, W. C., Westermarck, J. (2009). PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. Cancer Res. 69, 2870-2877.
9. Ortega-Gutierrez, S., Leung, D., Ficarro, S., Peters, E. C., Cravatt, B. F. (2008). Targeted disruption of the PME-1 gene causes loss of demethylated PP2A and perinatal lethality in mice. PLoS ONE 3, e2486.
10. Kidd, D., Liu, Y., Cravatt, B. F. (2001). Profiling serine hydrolase activities in complex proteomes. Biochemistry 40, 4005-4015.

Keywords:

Summary, Summary AID, PME-1, protein phosphatase methylesterase 1, PPME-1, protein phosphatase 2a, PP2a, lysophospholipase, LYPLA1, LYPLA2, cancer, fluorescence polarization, activity-based protein profiling, ABPP, fluorophosphonate rhodamine, FP-Rh, antagonist, inhibitor, counterscreen, high throughput screen, HTS, 1536, Scripps, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Panel Information
AIDs
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1AID 493154Abhydrolase domain containing 11 [Mus musculus] [gi:47682716]
Taxonomy id: 10090
Gene id: 68758
2AID 493110_1Lypla1 [Mus musculus] [gi:71059731]
Taxonomy id: 10090
Gene id: 18777
3AID 493110_2lysophospholipase 2 [Mus musculus] [gi:123122209]
Taxonomy id: 10090
Gene id: 26394
4AID 493161_1cytotoxicity, serum-free media
5AID 493161_2cytotoxicity, media plus FCS
6AID 504482Abhydrolase domain containing 11 [Mus musculus] [gi:47682716]
Taxonomy id: 10090
Gene id: 68758
7AID 504507Abhydrolase domain containing 11 [Mus musculus] [gi:47682716]
Taxonomy id: 10090
Gene id: 68758
8AID 504510_1cytotoxicity, serum-free media
9AID 504510_2cytotoxicity, media plus FCS

§ Panel component ID.
Protocol
Details of protocols, compound structures, and results from the original assays can be found in PubChem at the respective AIDs.
Comment
A probe development project is underway at the SRIMSC.
Result Definitions
Show more
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 493154]*Data from AID 493154. The value for concentration at which 50% inhibition is observed; IC50 shown in uM.1Abhydrolase domain containing 11 [Mus musculus]FloatμM
3IC50 Qualifier [AID 493110_1]Data from AID 493110, Panel 1. Qualifier identifies if the resultant fold selectivity was determined to be less than or greater than its listed fold selectivity.2Lypla1 [Mus musculus]String
4IC50 [AID 493110_1]*Data from AID 493110, Panel 1. The value for concentration at which 50% inhibition is observed; IC50 shown in uM.2FloatμM
5IC50 Qualifier [AID 493110_2]Data from AID 493110, Panel 2. Qualifier identifies if the resultant fold selectivity was determined to be less than or greater than its listed fold selectivity.3lysophospholipase 2 [Mus musculus]String
6IC50 [AID 493110_2]*Data from AID 493110, Panel 2. The value for concentration at which 50% inhibition is observed; IC50 shown in uM.3FloatμM
7IC50 Qualifier [AID 493161_1]Data from AID 493161, Panel 1. Qualifier identifies if the resultant fold selectivity was determined to be less than or greater than its listed fold selectivity.4String
8IC50 [AID 493161_1]*Data from AID 493161, Panel 1. The value for concentration at which 50% inhibition is observed; CC50 shown in uM.4FloatμM
9IC50 Qualifier [AID 493161_2]Data from AID 493161, Panel 2. Qualifier identifies if the resultant fold selectivity was determined to be less than or greater than its listed fold selectivity.5String
10IC50 [AID 493161_2]*Data from AID 493161, Panel 2. The value for concentration at which 50% inhibition is observed; CC50 shown in uM.5FloatμM
11IC50 [AID 504482]*Data from AID 504482. The value for concentration at which 50% inhibition is observed in situ; IC50 shown in micromolar.6Abhydrolase domain containing 11 [Mus musculus]FloatμM
12IC50 [AID 504507]*Data from AID 504507. The value for concentration at which 50% inhibition is observed; IC50 shown in uM.7Abhydrolase domain containing 11 [Mus musculus]FloatμM
13IC50 Qualifier [AID 504510_1]Data from AID 504510, Panel 1. 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.8String
14IC50 [AID 504510_1]*Data from AID 504510, Panel 1. The value for concentration at which 50% of surviving cells are observed; CC50 shown in micromolar.8FloatμM
15IC50 Qualifier [AID 504510_2]Data from AID 504510, 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.9String
16IC50 [AID 504510_2]*Data from AID 504510, Panel 2. The value for concentration at which 50% of surviving cells are observed; CC50 shown in micromolar.9FloatμM

* Activity Concentration. § Panel component ID.
Additional Information
Grant Number: 1 R01 CA132630

Classification
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