Name: 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. ..more
Target
| Sequence: monoacylglycerol lipase ABHD6 [Mus musculus] Protein Family: Lipase Gene:ABHD6 Related Protein 3D Structures |
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 | Confirmation screen (DAGLB inhibitors in triplicate) | |
| 602311 | 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 serine hydrolases in mouse brain membrane in vitro set 1 | other | Late stage assay (serine hydrolases in mouse brain membrane inhibitors in vitro) | |
| 602312 | 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 anti-target ABHD11 in vitro set 2 | other | Late stage assay (ABHD11 inhibitors in vitro) | |
| 602319 | 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 antitarget ABHD6 in vitro | other | Late stage assay (ABHD6 inhibitors in vitro) | |
| 602320 | 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 DAGLb in vitro | confirmatory | Late stage assay (DAGLb inhibitors in vitro in triplicate) | |
| 602321 | 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 serine hydrolases in mouse brain membrane in vitro set 3 | other | Late stage assay (serine hydrolases in mouse brain membrane inhibitors in vitro) | |
| 602322 | 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 | confirmatory | Late stage assay (ABHD6 inhibitors in vitro in triplicate) | |
| 602323 | 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 serine hydrolases in mouse brain membrane in vitro set 2 | other | Late stage assay (serine hydrolases in mouse brain membrane inhibitors in vitro) | |
| 602299 | 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 overexpressed DAGLb in vitro set 2 | other | Late stage assay (overexpressed DAGLb inhibitors in vitro) | |
| 602300 | 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 overexpressed DAGLb in vitro set 3 | other | Late stage assay (overexpressed DAGLb inhibitors in vitro) | |
| 602301 | 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 anti-target ABHD11 in vitro set 1 | other | Late stage assay (ABHD11 inhibitors) | |
| 602302 | 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 overexpressed DAGLb in vitro set 1 | other | Late stage assay (overexpressed DAGLb inhibitors in vitro) | |
| 602303 | 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 overexpressed DAGLb in vitro; triazole urea library | other | Late stage assay (overexpressed DAGLb inhibitors in vitro; triazole urea library) | |
| 602335 | 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 anti-target ABHD6 in situ | confirmatory | Late stage assay (ABHD6 inhibitors in situ in triplicate) | |
| 602337 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): absorbance-based cell-based dose response assay to determine cytotoxicity of inhibitor compounds | confirmatory | Late stage dose response assay (cytotoxicity in five replicates) | |
| 602339 | 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 | other | Late stage assay (DAGLb SILAC selectivity analysis) | |
| 602341 | 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 for ABHD6 | other | Late stage assay (ABHD6 SILAC selectivity analysis) | |
| 602343 | 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 brain ABHD6 in vivo, set 2 | other | Late stage assay (mouse brain ABHD6 inhibitors in vivo) | |
| 602345 | 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 macrophage DAGLb in vivo | other | Late stage assay (mouse macrophage DAGLb inhibitors in vivo in duplicate) | |
| 602347 | 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 brain ABHD6 in vivo | other | Late stage assay (mouse brain ABHD6 inhibitors in vivo in duplicate) | |
| 602349 | Late stage assay provider results from the probe development effort to identify inhibitors of DAGLb: fluorescence-based biochemical gel-based competitive Activity-Based Protein Profiling (ABPP) assay to distinguish systemic and peripheral inhibitors | other | Late stage assay (distinguish systemic and peripheral inhibitors in duplicate) | |
| 602351 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based animal-based Activity-Based Protein Profiling (ABPP) MudPIT selectivity analysis for ABHD6 | other | Late stage assay (ABHD6 MudPIT selectivity analysis in triplicate) | |
| 602353 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): LCMS-based animal-based Activity-Based Protein Profiling (ABPP) MudPIT selectivity analysis | other | Late stage assay (DAGLb MudPIT selectivity analysis in triplicate) | |
| 602354 | Late stage assay provider results from the probe development effort to identify inhibitors of diacylglycerol lipase, beta (DAGLb): fluorescence-based biochemical dose-response gel-based competitive Activity-Based Protein Profiling (ABPP) inhibition of DAGLb in situ | confirmatory | Late stage dose-response (DAGLb inhibitors in situ in triplicate) | |
| 602355 | 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 serine hydrolases in vitro | other | Late stage assay (serine hydrolases inhibitors in vitro) | |
| 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 | Late stage assay (recombinant DAGLa inhibitors in vitro) | |
| 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 | Late stage assay (DAGLb substrate turnover inhibitors in vitro) | |
| 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 | ||
| 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 | ||
| 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 |
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: ABHD6_INH_FLUO_3XIC50_INVITRO_SET2
Name: 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.
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, abhydrolase domain containing protein 6, ABHD6, activity-based protein profiling, ABPP, gel-based, activity-based probe, HT-01, inhibitor, inhibition, dose response, IC50, 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: ABHD6_INH_FLUO_3XIC50_INVITRO_SET2
Name: 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.
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, abhydrolase domain containing protein 6, ABHD6, activity-based protein profiling, ABPP, gel-based, activity-based probe, HT-01, inhibitor, inhibition, dose response, IC50, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Protocol
Assay Overview:
The purpose of this assay is to determine the IC50 values of powder samples of test compounds for anti-target ABHD6 inhibition in a complex proteome lysate. In this assay, a fluorescently-labeled activity-based probe, HT-01, which selectively labels several serine hydrolases including ABHD6, is used to label the proteins in the presence of test compounds. The reaction products are separated by SDS-PAGE and visualized in-gel using a flatbed fluorescence scanner. The percentage activity remaining is determined by measuring the integrated optical density (IOD) of the bands. As designed, test compounds that act as ABHD6 inhibitors will prevent enzyme-probe interactions, thereby decreasing the proportion of bound fluorescent probe, giving lower fluorescence intensity in the band in the gel.
Protocol Summary:
Membrane proteome of Neuro-2A murine neuroblastoma cells (25 uL of 1 mg/mL) in Dulbecco's PBS (DPBS) was treated with test compound (0.5 uL of a 50x stock in DMSO) or DMSO (0.5 uL) for 30 minutes at 37 C. The activity-based probe HT-01 (0.5 uL of a 50x stock in DMSO; 1 uM final concentration) was added, and the reaction was incubated for 30 minutes at 37 C, quenched with an equal volume of 2x SDS-PAGE loading buffer (reducing), separated by SDS-PAGE, and visualized by in-gel fluorescent scanning. The percentage activity remaining for anti-target ABHD6 was determined by measuring the integrated optical density of the individual protein bands relative to the DMSO-only (no compound) control. IC50 values were determined from dose-response curves from three replicates at each inhibitor concentration (6-point 1:5 dilution series from 2 uM to 0.00064 uM [2000 nM to 0.64 nM]).
The percent inhibition for each compound was calculated as follows:
%_Inhibition = ( 1 -( IOD_Test_Compound - Median_IOD_Low_Control ) / ( Median_IOD_High_Control - Median_IOD_Low_Control ) ) * 100
Where:
Test_Compound is defined as proteome treated with test compound.
High_Control is defined as proteome treated with DMSO only (no compound).
Low_Control is defined as background in a blank region of the gel.
Percent inhibition was plotted against the log of the compound concentration. A three parameter equation describing a sigmoidal dose-response curve was then fitted using GraphPad Prism (GraphPad Software Inc). The software-generated IC50 values are reported. In the event that Prism did not adequately fit the data, the IC50 is manually determined and reported as an approximate value.
PubChem Activity Outcome and Score:
Compounds with an IC50 less than or equal to 0.005 uM were considered active. Compounds with an IC50 greater than 0.005 uM were considered inactive.
Activity score was then ranked by the potency, with the most potent compounds assigned the highest activity scores.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A membrane proteome (provided by Assay Provider)
HT-01 (provided by Assay Provider)
DPBS (Cellgro 20-031-CV)
The purpose of this assay is to determine the IC50 values of powder samples of test compounds for anti-target ABHD6 inhibition in a complex proteome lysate. In this assay, a fluorescently-labeled activity-based probe, HT-01, which selectively labels several serine hydrolases including ABHD6, is used to label the proteins in the presence of test compounds. The reaction products are separated by SDS-PAGE and visualized in-gel using a flatbed fluorescence scanner. The percentage activity remaining is determined by measuring the integrated optical density (IOD) of the bands. As designed, test compounds that act as ABHD6 inhibitors will prevent enzyme-probe interactions, thereby decreasing the proportion of bound fluorescent probe, giving lower fluorescence intensity in the band in the gel.
Protocol Summary:
Membrane proteome of Neuro-2A murine neuroblastoma cells (25 uL of 1 mg/mL) in Dulbecco's PBS (DPBS) was treated with test compound (0.5 uL of a 50x stock in DMSO) or DMSO (0.5 uL) for 30 minutes at 37 C. The activity-based probe HT-01 (0.5 uL of a 50x stock in DMSO; 1 uM final concentration) was added, and the reaction was incubated for 30 minutes at 37 C, quenched with an equal volume of 2x SDS-PAGE loading buffer (reducing), separated by SDS-PAGE, and visualized by in-gel fluorescent scanning. The percentage activity remaining for anti-target ABHD6 was determined by measuring the integrated optical density of the individual protein bands relative to the DMSO-only (no compound) control. IC50 values were determined from dose-response curves from three replicates at each inhibitor concentration (6-point 1:5 dilution series from 2 uM to 0.00064 uM [2000 nM to 0.64 nM]).
The percent inhibition for each compound was calculated as follows:
%_Inhibition = ( 1 -( IOD_Test_Compound - Median_IOD_Low_Control ) / ( Median_IOD_High_Control - Median_IOD_Low_Control ) ) * 100
Where:
Test_Compound is defined as proteome treated with test compound.
High_Control is defined as proteome treated with DMSO only (no compound).
Low_Control is defined as background in a blank region of the gel.
Percent inhibition was plotted against the log of the compound concentration. A three parameter equation describing a sigmoidal dose-response curve was then fitted using GraphPad Prism (GraphPad Software Inc). The software-generated IC50 values are reported. In the event that Prism did not adequately fit the data, the IC50 is manually determined and reported as an approximate value.
PubChem Activity Outcome and Score:
Compounds with an IC50 less than or equal to 0.005 uM were considered active. Compounds with an IC50 greater than 0.005 uM were considered inactive.
Activity score was then ranked by the potency, with the most potent compounds assigned the highest activity scores.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
Neuro-2A membrane proteome (provided by Assay Provider)
HT-01 (provided by Assay Provider)
DPBS (Cellgro 20-031-CV)
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 | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||
| Score | The BioAssay activity ranking score |  | Integer | |||
| 1 | Qualifier | Indicates that the approximate IC50 value was manually determined. | String | |||
| 2 | Average IC50* | The average concentration at which 50 percent of the activity in the antagonist assay is observed; (IC50) shown in micromolar. | | Float | μM | |
| 3 | Inhibition at 2 uM [1] (2μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 2 uM compound; replicate [1]. | | Integer | % | |
| 4 | Inhibition at 0.4 uM [1] (0.4μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.400 uM compound; replicate [1]. | | Integer | % | |
| 5 | Inhibition at 0.08 uM [1] (0.08μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.080 uM compound; replicate [1]. | | Integer | % | |
| 6 | Inhibition at 0.016 uM [1] (0.016μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.016 uM compound; replicate [1]. | | Integer | % | |
| 7 | Inhibition at 0.0032 uM [1] (0.0032μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.0032 uM compound; replicate [1]. | | Integer | % | |
| 8 | Inhibition at 0.00064 uM [1] (0.00064μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.00064 uM compound; replicate [1]. | | Integer | % | |
| 9 | Inhibition at 2 uM [2] (2μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 2 uM compound; replicate [2]. | | Integer | % | |
| 10 | Inhibition at 0.4 uM [2] (0.4μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.400 uM compound; replicate [2]. | | Integer | % | |
| 11 | Inhibition at 0.08 uM [2] (0.08μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.080 uM compound; replicate [2]. | | Integer | % | |
| 12 | Inhibition at 0.016 uM [2] (0.016μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.016 uM compound; replicate [2]. | | Integer | % | |
| 13 | Inhibition at 0.0032 uM [2] (0.0032μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.0032 uM compound; replicate [2]. | | Integer | % | |
| 14 | Inhibition at 0.00064 uM [2] (0.00064μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.00064 uM compound; replicate [2]. | | Integer | % | |
| 15 | Inhibition at 2 uM [3] (2μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 2 uM compound; replicate [3]. | | Integer | % | |
| 16 | Inhibition at 0.4 uM [3] (0.4μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.400 uM compound; replicate [3]. | | Integer | % | |
| 17 | Inhibition at 0.08 uM [3] (0.08μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.080 uM compound; replicate [3]. | | Integer | % | |
| 18 | Inhibition at 0.016 uM [3] (0.016μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.016 uM compound; replicate [3]. | | Integer | % | |
| 19 | Inhibition at 0.0032 uM [3] (0.0032μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.0032 uM compound; replicate [3]. | | Integer | % | |
| 20 | Inhibition at 0.00064 uM [3] (0.00064μM**) | The value for percent inhibition of endogenous mouse ABHD6 at 0.00064 uM compound; replicate [3]. | | Integer | % |
* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1 R01 DA025285
Data Table (Concise)
Classification
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