Name: Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis. ..more
BioActive Compound: 1
Depositor Specified Assays
Show more |
| AID | Name | Type | Probe | Comment |
|---|---|---|---|---|
| 504411 | Fluorescence-based primary biochemical high throughput screening assay to identify inhibitors of human diacylglycerol lipase, beta (DAGLB) | screening | Primary screen (DAGLB inhibitors in singlicate) | |
| 504420 | Summary of the probe development efforts to identify inhibitors of human diacylglycerol lipase, beta (DAGLB) | summary | 3 | Summary (DAGLB inhibitors) |
| 504445 | Fluorescence-based biochemical high throughput confirmation assay for inhibitors of human diacylglycerol lipase, beta (DAGLB) | screening | ABPP screen (DAGLb inhibitors in triplicate) | |
| 624039 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based dose-response biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of antitarget ABHD6 in vitro, set 2 | confirmatory | ||
| 624077 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse liver ABHD6 in vivo upon oral compound administration | other | ||
| 602415 | Assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LC/MS-based biochemical inhibition of overexpressed DAGLb substrate turnover in vitro | other | ||
| 624468 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based biochemical dose response assay | confirmatory | ||
| 624472 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of DAGLb by enantiomers of KT116 | other | ||
| 602403 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of recombinant antitarget DAGLa in vitro | other | ||
| 624041 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of mouse liver ABHD6 in vivo | other |
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Benjamin Cravatt, The Scripps Research Institute (TSRI)
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 DA025285
Grant Proposal PI: Benjamin Cravatt, The Scripps Research Institute (TSRI)
External Assay ID: DAGLB_INH_LCMS_ABPPSILAC
Name: Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis.
Description:
Endocannabinoids (ECs) represent a unique group of lipids that function as chemical messengers in the nervous system. To date, the two principle ECs identified in mammals are N-arachidonoyl-ethanolamine (anandamide) and 2-arachidonoyl-glycerol (2-AG). They have been implicated in various physiological and pathological functions including appetite, pain, sensation, memory, and addiction (1). Unlike traditional neurotransmitters, which are stored in vesicles, ECs are synthesized and released on demand, and then rapidly degraded to terminate signaling. Thus, the metabolic pathways that govern EC turnover are critical in determining the magnitude and duration of neuronal signaling events (2). Endocannabinoid biosynthesis, in contrast to degradation, is poorly understood. Recently, two serine hydrolases, DAGL-a and -B, were cloned and found to selectively cleave sn-1 acyl chains from diacylglycerols (DAG) to generate 2-AG in vitro (3). Their function in the nervous system was validated in vivo by the generation of DAGL-a and -B knock-out mice (4, 5). However, it is still unclear to what extent DAGL-a/B catalytic activity contributes to 2-AG-mediated signaling. The development of potent and selective inhibitors would offer a means to perturb DAGL-a/B activity in a selective, reversible, and temporally-controlled manner. Given the non-selective nature of current DAGL-a/B inhibitors (6), specific chemical probes would serve as invaluable tools to delineate DAGL-a/B function in 2-AG signaling networks of the brain.
References:
1. Di Marzo, V. (2008) Targeting the endocannabinoid system: to enhance or reduce?, Nat Rev Drug Discov 7, 438-455.
2. Ahn, K., McKinney, M. K., and Cravatt, B. F. (2008) Enzymatic pathways that regulate endocannabinoid signaling in the nervous system, Chem Rev 108, 1687-1707.
3. Bisogno, T., Howell, F., Williams, G., Minassi, A., Cascio, M. G., Ligresti, A., Matias, I., Schiano-Moriello, A., Paul, P., Williams, E. J., Gangadharan, U., Hobbs, C., Di Marzo, V., and Doherty, P. (2003) Cloning of the first sn1-DAG lipases points to the spatial and temporal regulation of endocannabinoid signaling in the brain, J Cell Biol 163, 463-468.
4. Gao, Y., Vasilyev, D. V., Goncalves, M. B., Howell, F. V., Hobbs, C., Reisenberg, M., Shen, R., Zhang, M. Y., Strassle, B. W., Lu, P., Mark, L., Piesla, M. J., Deng, K., Kouranova, E. V., Ring, R. H., Whiteside, G. T., Bates, B., Walsh, F. S., Williams, G., Pangalos, M. N., Samad, T. A., and Doherty, P. (2010) Loss of Retrograde Endocannabinoid Signaling and Reduced Adult Neurogenesis in Diacylglycerol Lipase Knock-out Mice, J Neurosci 30, 2017-2024.
5. Tanimura, A., Yamazaki, M., Hashimotodani, Y., Uchigashima, M., Kawata, S., Abe, M., Kita, Y., Hashimoto, K., Shimizu, T., Watanabe, M., Sakimura, K., and Kano, M. (2010) The Endocannabinoid 2-Arachidonoylglycerol Produced by Diacylglycerol Lipase +/- Mediates Retrograde Suppression of Synaptic Transmission, Neuron 65, 320-327.
6. Hoover, H. S., Blankman, J. L., Niessen, S., and Cravatt, B. F. (2008) Selectivity of inhibitors of endocannabinoid biosynthesis evaluated by activity-based protein profiling, Bioorganic & Medicinal Chemistry Letters 18, 5838-5841.
Keywords:
late stage, late stage AID, assay provider, powders, counterscreen, diacylglycerol lipase, diacylglycerol lipase-beta, DAGL, DAGL-beta, DAGLB, hydrolase, serine hydrolase, appetite, pain, sensation, memory, addiction, liquid chromatography, LC, tandem mass spectrometry, MS/MS, activity-based protein profiling, ABPP, stable isotope labeling with amino acids in cell culture, SILAC, ABPP-SILAC, inhibitor, inhibition, fluorophosphonate-biotin, FP-biotin, in situ, Neuro-2A, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Benjamin Cravatt, The Scripps Research Institute (TSRI)
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 DA025285
Grant Proposal PI: Benjamin Cravatt, The Scripps Research Institute (TSRI)
External Assay ID: DAGLB_INH_LCMS_ABPPSILAC
Name: Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based cell-based Activity-Based Protein Profiling (ABPP) SILAC selectivity analysis.
Description:
Endocannabinoids (ECs) represent a unique group of lipids that function as chemical messengers in the nervous system. To date, the two principle ECs identified in mammals are N-arachidonoyl-ethanolamine (anandamide) and 2-arachidonoyl-glycerol (2-AG). They have been implicated in various physiological and pathological functions including appetite, pain, sensation, memory, and addiction (1). Unlike traditional neurotransmitters, which are stored in vesicles, ECs are synthesized and released on demand, and then rapidly degraded to terminate signaling. Thus, the metabolic pathways that govern EC turnover are critical in determining the magnitude and duration of neuronal signaling events (2). Endocannabinoid biosynthesis, in contrast to degradation, is poorly understood. Recently, two serine hydrolases, DAGL-a and -B, were cloned and found to selectively cleave sn-1 acyl chains from diacylglycerols (DAG) to generate 2-AG in vitro (3). Their function in the nervous system was validated in vivo by the generation of DAGL-a and -B knock-out mice (4, 5). However, it is still unclear to what extent DAGL-a/B catalytic activity contributes to 2-AG-mediated signaling. The development of potent and selective inhibitors would offer a means to perturb DAGL-a/B activity in a selective, reversible, and temporally-controlled manner. Given the non-selective nature of current DAGL-a/B inhibitors (6), specific chemical probes would serve as invaluable tools to delineate DAGL-a/B function in 2-AG signaling networks of the brain.
References:
1. Di Marzo, V. (2008) Targeting the endocannabinoid system: to enhance or reduce?, Nat Rev Drug Discov 7, 438-455.
2. Ahn, K., McKinney, M. K., and Cravatt, B. F. (2008) Enzymatic pathways that regulate endocannabinoid signaling in the nervous system, Chem Rev 108, 1687-1707.
3. Bisogno, T., Howell, F., Williams, G., Minassi, A., Cascio, M. G., Ligresti, A., Matias, I., Schiano-Moriello, A., Paul, P., Williams, E. J., Gangadharan, U., Hobbs, C., Di Marzo, V., and Doherty, P. (2003) Cloning of the first sn1-DAG lipases points to the spatial and temporal regulation of endocannabinoid signaling in the brain, J Cell Biol 163, 463-468.
4. Gao, Y., Vasilyev, D. V., Goncalves, M. B., Howell, F. V., Hobbs, C., Reisenberg, M., Shen, R., Zhang, M. Y., Strassle, B. W., Lu, P., Mark, L., Piesla, M. J., Deng, K., Kouranova, E. V., Ring, R. H., Whiteside, G. T., Bates, B., Walsh, F. S., Williams, G., Pangalos, M. N., Samad, T. A., and Doherty, P. (2010) Loss of Retrograde Endocannabinoid Signaling and Reduced Adult Neurogenesis in Diacylglycerol Lipase Knock-out Mice, J Neurosci 30, 2017-2024.
5. Tanimura, A., Yamazaki, M., Hashimotodani, Y., Uchigashima, M., Kawata, S., Abe, M., Kita, Y., Hashimoto, K., Shimizu, T., Watanabe, M., Sakimura, K., and Kano, M. (2010) The Endocannabinoid 2-Arachidonoylglycerol Produced by Diacylglycerol Lipase +/- Mediates Retrograde Suppression of Synaptic Transmission, Neuron 65, 320-327.
6. Hoover, H. S., Blankman, J. L., Niessen, S., and Cravatt, B. F. (2008) Selectivity of inhibitors of endocannabinoid biosynthesis evaluated by activity-based protein profiling, Bioorganic & Medicinal Chemistry Letters 18, 5838-5841.
Keywords:
late stage, late stage AID, assay provider, powders, counterscreen, diacylglycerol lipase, diacylglycerol lipase-beta, DAGL, DAGL-beta, DAGLB, hydrolase, serine hydrolase, appetite, pain, sensation, memory, addiction, liquid chromatography, LC, tandem mass spectrometry, MS/MS, activity-based protein profiling, ABPP, stable isotope labeling with amino acids in cell culture, SILAC, ABPP-SILAC, inhibitor, inhibition, fluorophosphonate-biotin, FP-biotin, in situ, Neuro-2A, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Panel Information
Targets
§ Panel component ID.
| Data Table(Active) Data Table(All) | Show more |
| PID§ | Name | Substance | Panel Targets | Description | Additional Information | ||
|---|---|---|---|---|---|---|---|
| Active | Inactive | ||||||
| 1 | Plat | 1 | tissue-type plasminogen activator preproprotein [Mus musculus] [gi:170763477] | Taxonomy id: 10090 Gene id: 18791 | |||
| 2 | Lonp1 | 1 | lon protease homolog, mitochondrial precursor [Mus musculus] [gi:116089322] | Taxonomy id: 10090 Gene id: 74142 | |||
| 3 | Ctsa | 1 | lysosomal protective protein isoform b precursor [Mus musculus] [gi:84042523] | Taxonomy id: 10090 Gene id: 19025 | |||
| 4 | Lyplal1 | 1 | lysophospholipase-like protein 1 [Mus musculus] [gi:227496223] | Taxonomy id: 10090 Gene id: 226791 | |||
| 5 | Scpep1 | 1 | retinoid-inducible serine carboxypeptidase precursor [Mus musculus] [gi:253970508] | Taxonomy id: 10090 Gene id: 74617 | |||
| 6 | Qrsl1 | 1 | glutamyl-tRNA(Gln) amidotransferase subunit A, mitochondrial [Mus musculus] [gi:124486688] | Taxonomy id: 10090 Gene id: 76563 | |||
| 7 | Pnpla7 | 1 | patatin-like phospholipase domain-containing protein 7 [Mus musculus] [gi:225007615] | Taxonomy id: 10090 Gene id: 241274 | |||
| 8 | Abhd11 | 1 | abhydrolase domain-containing protein 11 isoform 1 [Mus musculus] [gi:21644577] | Taxonomy id: 10090 Gene id: 68758 | |||
| 9 | Ache | 1 | acetylcholinesterase precursor [Mus musculus] [gi:13928664] | Taxonomy id: 10090 Gene id: 11423 | |||
| 10 | Serhl | 1 | serine hydrolase-like protein [Mus musculus] [gi:13443008] | Taxonomy id: 10090 Gene id: 68607 | |||
| 11 | Ppt2 | 1 | lysosomal thioesterase PPT2 precursor [Mus musculus] [gi:9506985] | Taxonomy id: 10090 Gene id: 54397 | |||
| 12 | Acot2 | 1 | acyl-coenzyme A thioesterase 2, mitochondrial precursor [Mus musculus] [gi:238624114] | Taxonomy id: 10090 Gene id: 171210 | |||
| 13 | Bat5 | 1 | abhydrolase domain-containing protein 16A [Mus musculus] [gi:30519896] | Taxonomy id: 10090 Gene id: 193742 | |||
| 14 | Htra2 | 1 | fatty acid synthase [Mus musculus] [gi:93102409] | Taxonomy id: 10090 Gene id: 64704 | |||
| 15 | Kiaa1363 | 1 | neutral cholesterol ester hydrolase 1 [Mus musculus] [gi:30520239] | Taxonomy id: 10090 Gene id: 320024 | |||
| 16 | Pnpla8 | 1 | calcium-independent phospholipase A2-gamma [Mus musculus] [gi:118130807] | Taxonomy id: 10090 Gene id: 67452 | |||
| 17 | Pnpla6 | 1 | neuropathy target esterase isoform 1 [Mus musculus] [gi:390608665] | Taxonomy id: 10090 Gene id: 50767 | |||
| 18 | Fam108c | 1 | abhydrolase domain-containing protein FAM108C1 [Mus musculus] [gi:158186616] | Taxonomy id: 10090 Gene id: 70178 | |||
| 19 | D230012E17Rik | 1 | GPI inositol-deacylase [Mus musculus] [gi:254028203] | Taxonomy id: 10090 Gene id: 241062 | |||
| 20 | Dpp7 | 1 | dipeptidyl peptidase 2 precursor [Mus musculus] [gi:31981425] | Taxonomy id: 10090 Gene id: 83768 | |||
| 21 | Lactb | 1 | serine beta-lactamase-like protein LACTB, mitochondrial precursor [Mus musculus] [gi:13507666] | Taxonomy id: 10090 Gene id: 80907 | |||
| 22 | Fam108b | 1 | abhydrolase domain-containing protein FAM108B1 precursor [Mus musculus] [gi:38142456] | Taxonomy id: 10090 Gene id: 226016 | |||
| 23 | Abhd12 | 1 | monoacylglycerol lipase ABHD12 [Mus musculus] [gi:159110817] | Taxonomy id: 10090 Gene id: 76192 | |||
| 24 | Esd | 1 | S-formylglutathione hydrolase [Mus musculus] [gi:13937355] | Taxonomy id: 10090 Gene id: 13885 | |||
| 25 | Lipe | 1 | hormone-sensitive lipase isoform 2 [Mus musculus] [gi:87239972] | Taxonomy id: 10090 Gene id: 16890 | |||
| 26 | Fasn | 1 | fatty acid synthase [Mus musculus] [gi:93102409] | Taxonomy id: 10090 Gene id: 14104 | |||
| 27 | Pme1 | 1 | protein phosphatase methylesterase 1 [Mus musculus] [gi:30794138] | Taxonomy id: 10090 Gene id: 72590 | |||
| 28 | Parl | 1 | presenilins-associated rhomboid-like protein, mitochondrial precursor [Mus musculus] [gi:54261813] | Taxonomy id: 10090 Gene id: 381038 | |||
| 29 | Faah | 1 | fatty acid amide hydrolase [Mus musculus] [gi:123253900] | Taxonomy id: 10090 Gene id: 14073 | |||
| 30 | Iah1 | 1 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] [gi:27754071] | Taxonomy id: 10090 Gene id: 67732 | |||
| 31 | Pafah1b2 | 1 | platelet-activating factor acetylhydrolase IB subunit beta [Mus musculus] [gi:40254624] | Taxonomy id: 10090 Gene id: 18475 | |||
| 32 | Abhd10 | 1 | abhydrolase domain-containing protein 10, mitochondrial precursor [Mus musculus] [gi:269784760] | Taxonomy id: 10090 Gene id: 213012 | |||
| 33 | Lypla2 | 1 | acyl-protein thioesterase 2 [Mus musculus] [gi:7242156] | Taxonomy id: 10090 Gene id: 26394 | |||
| 34 | Prep | 1 | prolyl endopeptidase [Mus musculus] [gi:6755152] | Taxonomy id: 10090 Gene id: 19072 | |||
| 35 | Prepl | 1 | prolyl endopeptidase-like isoform a [Mus musculus] [gi:254939518] | Taxonomy id: 10090 Gene id: 213760 | |||
| 36 | Pafah1b3 | 1 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] [gi:6679201] | Taxonomy id: 10090 Gene id: 18476 | |||
| 37 | Ddhd1 | 1 | phospholipase DDHD1 isoform 2 [Mus musculus] [gi:111955224] | Taxonomy id: 10090 Gene id: 114874 | |||
| 38 | Pafah2 | 1 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] [gi:225579137] | Taxonomy id: 10090 Gene id: 100163 | |||
| 39 | Apeh | 1 | Acylpeptide hydrolase [Mus musculus] [gi:19343726] | Taxonomy id: 10090 Gene id: 235606 | |||
| 40 | Lypla3 | 1 | group XV phospholipase A2 precursor [Mus musculus] [gi:19527008] | Taxonomy id: 10090 Gene id: 192654 | |||
| 41 | Abhd4 | 1 | abhydrolase domain-containing protein 4 isoform 1 [Mus musculus] [gi:326937491] | Taxonomy id: 10090 Gene id: 105501 | |||
| 42 | Dpp9 | 1 | dipeptidyl peptidase 9 [Mus musculus] [gi:255003757] | Taxonomy id: 10090 Gene id: 224897 | |||
| 43 | Lypla1 | 1 | acyl-protein thioesterase 1 [Mus musculus] [gi:6678760] | Taxonomy id: 10090 Gene id: 18777 | |||
| 44 | Tpp2 | 1 | Tpp2 protein [Mus musculus] [gi:37194903] | Taxonomy id: 10090 Gene id: 22019 | |||
| 45 | Abhd6 | 1 | monoacylglycerol lipase ABHD6 [Mus musculus] [gi:31560264] | Taxonomy id: 10090 Gene id: 66082 | |||
| 46 | Daglb | 1 | sn1-specific diacylglycerol lipase beta [Mus musculus] [gi:31559956] | Taxonomy id: 10090 Gene id: 231871 | |||
§ Panel component ID.
Protocol
Assay Overview:
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using activity-based protein profiling (ABPP) in combination with stable isotope labeling with amino acids in cell culture (SILAC). In this assay, cultured Neuro-2A cells are metabolically labeled with heavy or light amino acids. Heavy and light cells are treated with test compound and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with the serine-hydrolase-specific activity-based fluorophosphonate-biotin (FP-biotin) affinity probe, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by multi-dimensional liquid chromatography tandem mass spectrometery LC/LC-MS/MS (MudPIT). Inhibition of target and anti-target activity is quantified by comparing intensities of light and heavy peptide peaks. As designed, compounds that act as inhibitors will block FP-biotin labeling, reducing enrichment in the inhibitor-treated (heavy) sample relative to the DMSO-treated (light) sample, giving a smaller heavy/light ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Sample Preparation. Neuro-2A murine neuroblastoma cells were initially grown for 10 passages in either light or heavy SILAC DMEM medium supplemented with 10% dialyzed FCS and 2 mM L-glutamine. Light medium was supplemented with 100 ug/mL L-arginine and 100 ug/mL L-lysine. Heavy medium was supplemented with 100 ug/mL [13C615N4]-L-Arginine and 100 ug/mL [13C615N2]-L-Lysine. Heavy cells (in 10 mL medium) were treated with test compound KT172 (SID125269120, CID53364485) (10 uL of a 1000x stock in DMSO; 25 nM final concentration) and light cells were treated with DMSO (10 uL) for 4 hours at 37 C. Cells were washed with DPBS (4x), harvested, and homogenized by sonication in DPBS. The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration for each fraction was adjusted to 2 mg/mL with DPBS. The light and heavy proteomes were labeled with the activity-based affinity probe FP-biotin (500 uL total reaction volume, 10 uM final concentration) for 2 hours at 25 C. After incubation, light and heavy proteomes were mixed in 1:1 ratio, and the membrane proteomes were additionally solubilized with 1% Triton-X100. Samples were desalted over PD10 columns (GE Healthcare) in DPBS, and biotinylated proteins enriched with streptavidin beads (50 uL beads; conditions: 1 hour, 25 C 0.5% SDS in DPBS). The beads were washed with 1% SDS in DPBS (1x), 6 M urea (1x), and DPBS (2x), then resuspended in in 6 M urea (150 uL), reduced with 5 mM TCEP for 20 minutes, and alkylated with 10 mM iodoacetamide for 30 minutes at 25 C in the dark. The urea concentration was reduced to 2 M with 2x volume DPBS. On-bead digestions were performed for 12 hours at 37 C with sequence-grade modified trypsin (Promega; 2 ug) in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% (v/v) formic acid and stored at -80 C prior to analysis.
LC-MS/MS analysis. Samples were analyzed by multidimensional liquid chromatography tandem mass spectrometry (MudPIT) using an Agilent 1200-series quaternary pump and Thermo Scientific LTQ-Orbitrap Velos ion trap mass spectrometer. Peptides were eluted in a 5-step MudPIT experiment using 0%, 25%, 50%, 80%, and 100% salt bumps of 500 mM aqueous ammonium acetate and data were collected in data-dependent acquisition mode with dynamic exclusion turned on (20 seconds, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 30 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.9.2; publicly available at http://fields.scripps.edu/downloads.php). MS2 spectra data were searched using the ProLuCID algorithm (publicly available at http://fields.scripps.edu/downloads.php) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. ProLucid searches allowed for static modification of cysteine residues (+57.02146 due to alkylation), methionine oxidation (+15.9949), mass shifts of labeled amino acids (+10.0083 R, +8.0142 K) and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0) using the --modstat, --mass, and --trypstat options (applies different statistical models for the analysis of high resolution masses, peptide digestion state, and methionine oxidation state respectively). Ratios of heavy/light (test compound/DMSO) peaks were calculated using in-house software and normalized at the peptide level to the average ratio of all non-serine hydrolase peptides. Reported ratios represent the mean of all unique, quantified peptides per protein and do not include peptides that were >3 standard deviations from the median peptide value. Proteins with less than three peptides per protein ID were not included in the analysis.
Ratio = Average( AUClight / AUCheavy ) calculated for all unique peptides
Where:
AUClight is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUCheavy is the area-under-the-curve for the heavy peptide pair from cells treated with DMSO.
PubChem Activity Outcome and Score:
The following applies to each panel in this assay:
A compound was considered active for a particular target/anti-target with a light/heavy ratio of less than or equal to 0.5. A compound was considered inactive for a specified target/anti-target with a light/heavy ratio of greater than 0.5.
Overall Outcome and Score:
A compound was considered active if it was active for DAGLb and active for fewer than two anti-target serine hydrolases tested.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A murine cells (provided by Assay Provider)
SILAC DMEM media (Thermo 89985)
dialyzed FCS (Gemini 100-108)
1x Glutamine (CellGro 25-005-CI)
L-Arginine (SigmaAldrich A6969)
L-Lysine (SigmaAldrich L9037)
[13C615N4]-L-Arginine (SigmaAldrich 608033)
[13C615N2]-L-Lysine (SigmaAldrich 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using activity-based protein profiling (ABPP) in combination with stable isotope labeling with amino acids in cell culture (SILAC). In this assay, cultured Neuro-2A cells are metabolically labeled with heavy or light amino acids. Heavy and light cells are treated with test compound and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with the serine-hydrolase-specific activity-based fluorophosphonate-biotin (FP-biotin) affinity probe, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by multi-dimensional liquid chromatography tandem mass spectrometery LC/LC-MS/MS (MudPIT). Inhibition of target and anti-target activity is quantified by comparing intensities of light and heavy peptide peaks. As designed, compounds that act as inhibitors will block FP-biotin labeling, reducing enrichment in the inhibitor-treated (heavy) sample relative to the DMSO-treated (light) sample, giving a smaller heavy/light ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Sample Preparation. Neuro-2A murine neuroblastoma cells were initially grown for 10 passages in either light or heavy SILAC DMEM medium supplemented with 10% dialyzed FCS and 2 mM L-glutamine. Light medium was supplemented with 100 ug/mL L-arginine and 100 ug/mL L-lysine. Heavy medium was supplemented with 100 ug/mL [13C615N4]-L-Arginine and 100 ug/mL [13C615N2]-L-Lysine. Heavy cells (in 10 mL medium) were treated with test compound KT172 (SID125269120, CID53364485) (10 uL of a 1000x stock in DMSO; 25 nM final concentration) and light cells were treated with DMSO (10 uL) for 4 hours at 37 C. Cells were washed with DPBS (4x), harvested, and homogenized by sonication in DPBS. The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration for each fraction was adjusted to 2 mg/mL with DPBS. The light and heavy proteomes were labeled with the activity-based affinity probe FP-biotin (500 uL total reaction volume, 10 uM final concentration) for 2 hours at 25 C. After incubation, light and heavy proteomes were mixed in 1:1 ratio, and the membrane proteomes were additionally solubilized with 1% Triton-X100. Samples were desalted over PD10 columns (GE Healthcare) in DPBS, and biotinylated proteins enriched with streptavidin beads (50 uL beads; conditions: 1 hour, 25 C 0.5% SDS in DPBS). The beads were washed with 1% SDS in DPBS (1x), 6 M urea (1x), and DPBS (2x), then resuspended in in 6 M urea (150 uL), reduced with 5 mM TCEP for 20 minutes, and alkylated with 10 mM iodoacetamide for 30 minutes at 25 C in the dark. The urea concentration was reduced to 2 M with 2x volume DPBS. On-bead digestions were performed for 12 hours at 37 C with sequence-grade modified trypsin (Promega; 2 ug) in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% (v/v) formic acid and stored at -80 C prior to analysis.
LC-MS/MS analysis. Samples were analyzed by multidimensional liquid chromatography tandem mass spectrometry (MudPIT) using an Agilent 1200-series quaternary pump and Thermo Scientific LTQ-Orbitrap Velos ion trap mass spectrometer. Peptides were eluted in a 5-step MudPIT experiment using 0%, 25%, 50%, 80%, and 100% salt bumps of 500 mM aqueous ammonium acetate and data were collected in data-dependent acquisition mode with dynamic exclusion turned on (20 seconds, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 30 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.9.2; publicly available at http://fields.scripps.edu/downloads.php). MS2 spectra data were searched using the ProLuCID algorithm (publicly available at http://fields.scripps.edu/downloads.php) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. ProLucid searches allowed for static modification of cysteine residues (+57.02146 due to alkylation), methionine oxidation (+15.9949), mass shifts of labeled amino acids (+10.0083 R, +8.0142 K) and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0) using the --modstat, --mass, and --trypstat options (applies different statistical models for the analysis of high resolution masses, peptide digestion state, and methionine oxidation state respectively). Ratios of heavy/light (test compound/DMSO) peaks were calculated using in-house software and normalized at the peptide level to the average ratio of all non-serine hydrolase peptides. Reported ratios represent the mean of all unique, quantified peptides per protein and do not include peptides that were >3 standard deviations from the median peptide value. Proteins with less than three peptides per protein ID were not included in the analysis.
Ratio = Average( AUClight / AUCheavy ) calculated for all unique peptides
Where:
AUClight is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUCheavy is the area-under-the-curve for the heavy peptide pair from cells treated with DMSO.
PubChem Activity Outcome and Score:
The following applies to each panel in this assay:
A compound was considered active for a particular target/anti-target with a light/heavy ratio of less than or equal to 0.5. A compound was considered inactive for a specified target/anti-target with a light/heavy ratio of greater than 0.5.
Overall Outcome and Score:
A compound was considered active if it was active for DAGLb and active for fewer than two anti-target serine hydrolases tested.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A murine cells (provided by Assay Provider)
SILAC DMEM media (Thermo 89985)
dialyzed FCS (Gemini 100-108)
1x Glutamine (CellGro 25-005-CI)
L-Arginine (SigmaAldrich A6969)
L-Lysine (SigmaAldrich L9037)
[13C615N4]-L-Arginine (SigmaAldrich 608033)
[13C615N2]-L-Lysine (SigmaAldrich 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
Comment
This assay was performed by the assay provider with powder samples of synthetic compounds.
Categorized Comment
BAO: version: 1.4b1090
BAO: bioassay specification: assay stage: secondary: counter screening
BAO: bioassay specification: assay biosafety level: bsl1
BAO: assay format: biochemical format: protein format: single protein format
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: meta target: molecular target: protein target: enzyme: generic hydrolase
BAO: meta target: biological process target: regulation of molecular function
BAO: meta target detail: binding reporter specification: interaction: protein-small molecule
BAO: detection technology: fluorescence: fluorescence intensity
BAO: bioassay specification: assay stage: secondary: counter screening
BAO: bioassay specification: assay biosafety level: bsl1
BAO: assay format: biochemical format: protein format: single protein format
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: meta target: molecular target: protein target: enzyme: generic hydrolase
BAO: meta target: biological process target: regulation of molecular function
BAO: meta target detail: binding reporter specification: interaction: protein-small molecule
BAO: detection technology: fluorescence: fluorescence intensity
Result Definitions
| Show more |
| TID | Name | Description | PID§ | Panel Targets | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||||
| Score | The BioAssay activity ranking score |  | Integer | |||||
| 1 | Outcome [Plat] | One of Active, Inactive, or Not Tested | 1 | tissue-type plasminogen activator preproprotein [Mus musculus] | | Outcome | ||
| 2 | Mean Ratio [Plat] | Calculated inhibitor-treated/DMSO ratio for Plat as mean for all unique peptides quantified | 1 | | Float | ratio | ||
| 3 | Standard Error [Plat] | Calculated standard error for mean | 1 | | Float | |||
| 4 | Outcome [Lonp1] | One of Active, Inactive, or Not Tested | 2 | lon protease homolog, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 5 | Mean Ratio [Lonp1] | Calculated inhibitor-treated/DMSO ratio for Lonp1 as mean for all unique peptides quantified | 2 | | Float | ratio | ||
| 6 | Standard Error [Lonp1] | Calculated standard error for mean | 2 | | Float | |||
| 7 | Outcome [Ctsa] | One of Active, Inactive, or Not Tested | 3 | lysosomal protective protein isoform b precursor [Mus musculus] | | Outcome | ||
| 8 | Mean Ratio [Ctsa] | Calculated inhibitor-treated/DMSO ratio for Ctsa as mean for all unique peptides quantified | 3 | | Float | ratio | ||
| 9 | Standard Error [Ctsa] | Calculated standard error for mean | 3 | | Float | |||
| 10 | Outcome [Lyplal1] | One of Active, Inactive, or Not Tested | 4 | lysophospholipase-like protein 1 [Mus musculus] | | Outcome | ||
| 11 | Mean Ratio [Lyplal1] | Calculated inhibitor-treated/DMSO ratio for Lyplal1 as mean for all unique peptides quantified | 4 | | Float | ratio | ||
| 12 | Standard Error [Lyplal1] | Calculated standard error for mean | 4 | | Float | |||
| 13 | Outcome [Scpep1] | One of Active, Inactive, or Not Tested | 5 | retinoid-inducible serine carboxypeptidase precursor [Mus musculus] | | Outcome | ||
| 14 | Mean Ratio [Scpep1] | Calculated inhibitor-treated/DMSO ratio for Scpep1 as mean for all unique peptides quantified | 5 | | Float | ratio | ||
| 15 | Standard Error [Scpep1] | Calculated standard error for mean | 5 | | Float | |||
| 16 | Outcome [Qrsl1] | One of Active, Inactive, or Not Tested | 6 | glutamyl-tRNA(Gln) amidotransferase subunit A, mitochondrial [Mus musculus] | | Outcome | ||
| 17 | Mean Ratio [Qrsl1] | Calculated inhibitor-treated/DMSO ratio for Qrsl1 as mean for all unique peptides quantified | 6 | | Float | ratio | ||
| 18 | Standard Error [Qrsl1] | Calculated standard error for mean | 6 | | Float | |||
| 19 | Outcome [Pnpla7] | One of Active, Inactive, or Not Tested | 7 | patatin-like phospholipase domain-containing protein 7 [Mus musculus] | | Outcome | ||
| 20 | Mean Ratio [Pnpla7] | Calculated inhibitor-treated/DMSO ratio for Pnpla7 as mean for all unique peptides quantified | 7 | | Float | ratio | ||
| 21 | Standard Error [Pnpla7] | Calculated standard error for mean | 7 | | Float | |||
| 22 | Outcome [Abhd11] | One of Active, Inactive, or Not Tested | 8 | abhydrolase domain-containing protein 11 isoform 1 [Mus musculus] | | Outcome | ||
| 23 | Mean Ratio [Abhd11] | Calculated inhibitor-treated/DMSO ratio for Abhd11 as mean for all unique peptides quantified | 8 | | Float | ratio | ||
| 24 | Standard Error [Abhd11] | Calculated standard error for mean | 8 | | Float | |||
| 25 | Outcome [Ache] | One of Active, Inactive, or Not Tested | 9 | acetylcholinesterase precursor [Mus musculus] | | Outcome | ||
| 26 | Mean Ratio [Ache] | Calculated inhibitor-treated/DMSO ratio for Ache as mean for all unique peptides quantified | 9 | | Float | ratio | ||
| 27 | Standard Error [Ache] | Calculated standard error for mean | 9 | | Float | |||
| 28 | Outcome [Serhl] | One of Active, Inactive, or Not Tested | 10 | serine hydrolase-like protein [Mus musculus] | | Outcome | ||
| 29 | Mean Ratio [Serhl] | Calculated inhibitor-treated/DMSO ratio for Serhl as mean for all unique peptides quantified | 10 | | Float | ratio | ||
| 30 | Standard Error [Serhl] | Calculated standard error for mean | 10 | | Float | |||
| 31 | Outcome [Ppt2] | One of Active, Inactive, or Not Tested | 11 | lysosomal thioesterase PPT2 precursor [Mus musculus] | | Outcome | ||
| 32 | Mean Ratio [Ppt2] | Calculated inhibitor-treated/DMSO ratio for Ppt2 as mean for all unique peptides quantified | 11 | | Float | ratio | ||
| 33 | Standard Error [Ppt2] | Calculated standard error for mean | 11 | | Float | |||
| 34 | Outcome [Acot2] | One of Active, Inactive, or Not Tested | 12 | acyl-coenzyme A thioesterase 2, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 35 | Mean Ratio [Acot2] | Calculated inhibitor-treated/DMSO ratio for Acot2 as mean for all unique peptides quantified | 12 | | Float | ratio | ||
| 36 | Standard Error [Acot2] | Calculated standard error for mean | 12 | | Float | |||
| 37 | Outcome [Bat5] | One of Active, Inactive, or Not Tested | 13 | abhydrolase domain-containing protein 16A [Mus musculus] | | Outcome | ||
| 38 | Mean Ratio [Bat5] | Calculated inhibitor-treated/DMSO ratio for Bat5 as mean for all unique peptides quantified | 13 | | Float | ratio | ||
| 39 | Standard Error [Bat5] | Calculated standard error for mean | 13 | | Float | |||
| 40 | Outcome [Htra2] | One of Active, Inactive, or Not Tested | 14 | fatty acid synthase [Mus musculus] | | Outcome | ||
| 41 | Mean Ratio [Htra2] | Calculated inhibitor-treated/DMSO ratio for HtrA2 as mean for all unique peptides quantified | 14 | | Float | ratio | ||
| 42 | Standard Error [Htra2] | Calculated standard error for mean | 14 | | Float | |||
| 43 | Outcome [Kiaa1363] | One of Active, Inactive, or Not Tested | 15 | neutral cholesterol ester hydrolase 1 [Mus musculus] | | Outcome | ||
| 44 | Mean Ratio [Kiaa1363] | Calculated inhibitor-treated/DMSO ratio for Kiaa1363 as mean for all unique peptides quantified | 15 | | Float | ratio | ||
| 45 | Standard Error [Kiaa1363] | Calculated standard error for mean | 15 | | Float | |||
| 46 | Outcome [Pnpla8] | One of Active, Inactive, or Not Tested | 16 | calcium-independent phospholipase A2-gamma [Mus musculus] | | Outcome | ||
| 47 | Mean Ratio [Pnpla8] | Calculated inhibitor-treated/DMSO ratio for Pnpla8 as mean for all unique peptides quantified | 16 | | Float | ratio | ||
| 48 | Standard Error [Pnpla8] | Calculated standard error for mean | 16 | | Float | |||
| 49 | Outcome [Pnpla6] | One of Active, Inactive, or Not Tested | 17 | neuropathy target esterase isoform 1 [Mus musculus] | | Outcome | ||
| 50 | Mean Ratio [Pnpla6] | Calculated inhibitor-treated/DMSO ratio for Pnpla6 as mean for all unique peptides quantified | 17 | | Float | ratio | ||
| 51 | Standard Error [Pnpla6] | Calculated standard error for mean | 17 | | Float | |||
| 52 | Outcome [Fam108c] | One of Active, Inactive, or Not Tested | 18 | abhydrolase domain-containing protein FAM108C1 [Mus musculus] | | Outcome | ||
| 53 | Mean Ratio [Fam108c] | Calculated inhibitor-treated/DMSO ratio for Fam108c as mean for all unique peptides quantified | 18 | | Float | ratio | ||
| 54 | Standard Error [Fam108c] | Calculated standard error for mean | 18 | | Float | |||
| 55 | Outcome [D230012E17Rik] | One of Active, Inactive, or Not Tested | 19 | GPI inositol-deacylase [Mus musculus] | | Outcome | ||
| 56 | Mean Ratio [D230012E17Rik] | Calculated inhibitor-treated/DMSO ratio for D230012E17Rik as mean for all unique peptides quantified | 19 | | Float | ratio | ||
| 57 | Standard Error [D230012E17Rik] | Calculated standard error for mean | 19 | | Float | |||
| 58 | Outcome [Dpp7] | One of Active, Inactive, or Not Tested | 20 | dipeptidyl peptidase 2 precursor [Mus musculus] | | Outcome | ||
| 59 | Mean Ratio [Dpp7] | Calculated inhibitor-treated/DMSO ratio for Dpp7 as mean for all unique peptides quantified | 20 | | Float | ratio | ||
| 60 | Standard Error [Dpp7] | Calculated standard error for mean | 20 | | Float | |||
| 61 | Outcome [Lactb] | One of Active, Inactive, or Not Tested | 21 | serine beta-lactamase-like protein LACTB, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 62 | Mean Ratio [Lactb] | Calculated inhibitor-treated/DMSO ratio for Lactb as mean for all unique peptides quantified | 21 | | Float | ratio | ||
| 63 | Standard Error [Lactb] | Calculated standard error for mean | 21 | | Float | |||
| 64 | Outcome [Fam108b] | One of Active, Inactive, or Not Tested | 22 | abhydrolase domain-containing protein FAM108B1 precursor [Mus musculus] | | Outcome | ||
| 65 | Mean Ratio [Fam108b] | Calculated inhibitor-treated/DMSO ratio for Fam108b as mean for all unique peptides quantified | 22 | | Float | ratio | ||
| 66 | Standard Error [Fam108b] | Calculated standard error for mean | 22 | | Float | |||
| 67 | Outcome [Abhd12] | One of Active, Inactive, or Not Tested | 23 | monoacylglycerol lipase ABHD12 [Mus musculus] | | Outcome | ||
| 68 | Mean Ratio [Abhd12] | Calculated inhibitor-treated/DMSO ratio for Abhd12 as mean for all unique peptides quantified | 23 | | Float | ratio | ||
| 69 | Standard Error [Abhd12] | Calculated standard error for mean | 23 | | Float | |||
| 70 | Outcome [Esd] | One of Active, Inactive, or Not Tested | 24 | S-formylglutathione hydrolase [Mus musculus] | | Outcome | ||
| 71 | Mean Ratio [Esd] | Calculated inhibitor-treated/DMSO ratio for Esd as mean for all unique peptides quantified | 24 | | Float | ratio | ||
| 72 | Standard Error [Esd] | Calculated standard error for mean | 24 | | Float | |||
| 73 | Outcome [Lipe] | One of Active, Inactive, or Not Tested | 25 | hormone-sensitive lipase isoform 2 [Mus musculus] | | Outcome | ||
| 74 | Mean Ratio [Lipe] | Calculated inhibitor-treated/DMSO ratio for Lipe as mean for all unique peptides quantified | 25 | | Float | ratio | ||
| 75 | Standard Error [Lipe] | Calculated standard error for mean | 25 | | Float | |||
| 76 | Outcome [Fasn] | One of Active, Inactive, or Not Tested | 26 | fatty acid synthase [Mus musculus] | | Outcome | ||
| 77 | Mean Ratio [Fasn] | Calculated inhibitor-treated/DMSO ratio for Fasn as mean for all unique peptides quantified | 26 | | Float | ratio | ||
| 78 | Standard Error [Fasn] | Calculated standard error for mean | 26 | | Float | |||
| 79 | Outcome [Pme1] | One of Active, Inactive, or Not Tested | 27 | protein phosphatase methylesterase 1 [Mus musculus] | | Outcome | ||
| 80 | Mean Ratio [Pme1] | Calculated inhibitor-treated/DMSO ratio for Pme1 as mean for all unique peptides quantified | 27 | | Float | ratio | ||
| 81 | Standard Error [Pme1] | Calculated standard error for mean | 27 | | Float | |||
| 82 | Outcome [Parl] | One of Active, Inactive, or Not Tested | 28 | presenilins-associated rhomboid-like protein, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 83 | Mean Ratio [Parl] | Calculated inhibitor-treated/DMSO ratio for Parl as mean for all unique peptides quantified | 28 | | Float | ratio | ||
| 84 | Standard Error [Parl] | Calculated standard error for mean | 28 | | Float | |||
| 85 | Outcome [Faah] | One of Active, Inactive, or Not Tested | 29 | fatty acid amide hydrolase [Mus musculus] | | Outcome | ||
| 86 | Mean Ratio [Faah] | Calculated inhibitor-treated/DMSO ratio for Faah as mean for all unique peptides quantified | 29 | | Float | ratio | ||
| 87 | Standard Error [Faah] | Calculated standard error for mean | 29 | | Float | |||
| 88 | Outcome [Iah1] | One of Active, Inactive, or Not Tested | 30 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] | | Outcome | ||
| 89 | Mean Ratio [Iah1] | Calculated inhibitor-treated/DMSO ratio for Iah1 as mean for all unique peptides quantified | 30 | | Float | ratio | ||
| 90 | Standard Error [Iah1] | Calculated standard error for mean | 30 | | Float | |||
| 91 | Outcome [Pafah1b2] | One of Active, Inactive, or Not Tested | 31 | platelet-activating factor acetylhydrolase IB subunit beta [Mus musculus] | | Outcome | ||
| 92 | Mean Ratio [Pafah1b2] | Calculated inhibitor-treated/DMSO ratio for Pafah1b2 as mean for all unique peptides quantified | 31 | | Float | ratio | ||
| 93 | Standard Error [Pafah1b2] | Calculated standard error for mean | 31 | | Float | |||
| 94 | Outcome [Abhd10] | One of Active, Inactive, or Not Tested | 32 | abhydrolase domain-containing protein 10, mitochondrial precursor [Mus musculus] | | Outcome | ||
| 95 | Mean Ratio [Abhd10] | Calculated inhibitor-treated/DMSO ratio for Abhd10 as mean for all unique peptides quantified | 32 | | Float | ratio | ||
| 96 | Standard Error [Abhd10] | Calculated standard error for mean | 32 | | Float | |||
| 97 | Outcome [Lypla2] | One of Active, Inactive, or Not Tested | 33 | acyl-protein thioesterase 2 [Mus musculus] | | Outcome | ||
| 98 | Mean Ratio [Lypla2] | Calculated inhibitor-treated/DMSO ratio for Lypla2 as mean for all unique peptides quantified | 33 | | Float | ratio | ||
| 99 | Standard Error [Lypla2] | Calculated standard error for mean | 33 | | Float | |||
| 100 | Outcome [Prep] | One of Active, Inactive, or Not Tested | 34 | prolyl endopeptidase [Mus musculus] | | Outcome | ||
| 101 | Mean Ratio [Prep] | Calculated inhibitor-treated/DMSO ratio for Prep as mean for all unique peptides quantified | 34 | | Float | ratio | ||
| 102 | Standard Error [Prep] | Calculated standard error for mean | 34 | | Float | |||
| 103 | Outcome [Prepl] | One of Active, Inactive, or Not Tested | 35 | prolyl endopeptidase-like isoform a [Mus musculus] | | Outcome | ||
| 104 | Mean Ratio [Prepl] | Calculated inhibitor-treated/DMSO ratio for Prepl as mean for all unique peptides quantified | 35 | | Float | ratio | ||
| 105 | Standard Error [Prepl] | Calculated standard error for mean | 35 | | Float | |||
| 106 | Outcome [Pafah1b3] | One of Active, Inactive, or Not Tested | 36 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] | | Outcome | ||
| 107 | Mean Ratio [Pafah1b3] | Calculated inhibitor-treated/DMSO ratio for Pafah1b3 as mean for all unique peptides quantified | 36 | | Float | ratio | ||
| 108 | Standard Error [Pafah1b3] | Calculated standard error for mean | 36 | | Float | |||
| 109 | Outcome [Ddhd1] | One of Active, Inactive, or Not Tested | 37 | phospholipase DDHD1 isoform 2 [Mus musculus] | | Outcome | ||
| 110 | Mean Ratio [Ddhd1] | Calculated inhibitor-treated/DMSO ratio for Ddhd1 as mean for all unique peptides quantified | 37 | | Float | ratio | ||
| 111 | Standard Error [Ddhd1] | Calculated standard error for mean | 37 | | Float | |||
| 112 | Outcome [Pafah2] | One of Active, Inactive, or Not Tested | 38 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] | | Outcome | ||
| 113 | Mean Ratio [Pafah2] | Calculated inhibitor-treated/DMSO ratio for Pafah2 as mean for all unique peptides quantified | 38 | | Float | ratio | ||
| 114 | Standard Error [Pafah2] | Calculated standard error for mean | 38 | | Float | |||
| 115 | Outcome [Apeh] | One of Active, Inactive, or Not Tested | 39 | Acylpeptide hydrolase [Mus musculus] | | Outcome | ||
| 116 | Mean Ratio [Apeh] | Calculated inhibitor-treated/DMSO ratio for Apeh as mean for all unique peptides quantified | 39 | | Float | ratio | ||
| 117 | Standard Error [Apeh] | Calculated standard error for mean | 39 | | Float | |||
| 118 | Outcome [Lypla3] | One of Active, Inactive, or Not Tested | 40 | group XV phospholipase A2 precursor [Mus musculus] | | Outcome | ||
| 119 | Mean Ratio [Lypla3] | Calculated inhibitor-treated/DMSO ratio for Lypla3 as mean for all unique peptides quantified | 40 | | Float | ratio | ||
| 120 | Standard Error [Lypla3] | Calculated standard error for mean | 40 | | Float | |||
| 121 | Outcome [Abhd4] | One of Active, Inactive, or Not Tested | 41 | abhydrolase domain-containing protein 4 isoform 1 [Mus musculus] | | Outcome | ||
| 122 | Mean Ratio [Abhd4] | Calculated inhibitor-treated/DMSO ratio for Abhd4 as mean for all unique peptides quantified | 41 | | Float | ratio | ||
| 123 | Standard Error [Abhd4] | Calculated standard error for mean | 41 | | Float | |||
| 124 | Outcome [Dpp9] | One of Active, Inactive, or Not Tested | 42 | dipeptidyl peptidase 9 [Mus musculus] | | Outcome | ||
| 125 | Mean Ratio [Dpp9] | Calculated inhibitor-treated/DMSO ratio for Dpp9 as mean for all unique peptides quantified | 42 | | Float | ratio | ||
| 126 | Standard Error [Dpp9] | Calculated standard error for mean | 42 | | Float | |||
| 127 | Outcome [Lypla1] | One of Active, Inactive, or Not Tested | 43 | acyl-protein thioesterase 1 [Mus musculus] | | Outcome | ||
| 128 | Mean Ratio [Lypla1] | Calculated inhibitor-treated/DMSO ratio for Lypla1 as mean for all unique peptides quantified | 43 | | Float | ratio | ||
| 129 | Standard Error [Lypla1] | Calculated standard error for mean | 43 | | Float | |||
| 130 | Outcome [Tpp2] | One of Active, Inactive, or Not Tested | 44 | Tpp2 protein [Mus musculus] | | Outcome | ||
| 131 | Mean Ratio [Tpp2] | Calculated inhibitor-treated/DMSO ratio for Tpp2 as mean for all unique peptides quantified | 44 | | Float | ratio | ||
| 132 | Standard Error [Tpp2] | Calculated standard error for mean | 44 | | Float | |||
| 133 | Outcome [Abhd6] | One of Active, Inactive, or Not Tested | 45 | monoacylglycerol lipase ABHD6 [Mus musculus] | | Outcome | ||
| 134 | Mean Ratio [Abhd6] | Calculated inhibitor-treated/DMSO ratio for Abhd6 as mean for all unique peptides quantified | 45 | | Float | ratio | ||
| 135 | Standard Error [Abhd6] | Calculated standard error for mean | 45 | | Float | |||
| 136 | Outcome [Daglb] | One of Active, Inactive, or Not Tested | 46 | sn1-specific diacylglycerol lipase beta [Mus musculus] | | Outcome | ||
| 137 | Mean Ratio [Daglb] | Calculated inhibitor-treated/DMSO ratio for Daglb as mean for all unique peptides quantified | 46 | | Float | ratio | ||
| 138 | Standard Error [Daglb] | Calculated standard error for mean | 46 | | Float |
Additional Information
Grant Number: 1 R01 DA025285
Classification
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