Name: Late 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 ABHD11. ..more
BioActive Compound: 1
Depositor Specified Assays
Show more |
| AID | Name | Type | Probe | Comment |
|---|---|---|---|---|
| 2202 | Summary of probe development efforts to identify inhibitors of lysophospholipase 1 (LYPLA1). | summary | 2 | Summary (LYPLA1 inhibitors) |
| 2203 | Summary of probe development efforts to identify inhibitors of lysophospholipase 2 (LYPLA2). | summary | 1 | Summary (LYPLA2 inhibitors) |
| 2174 | Counterscreen for PME1 inhibitors: fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of lysophospholipase 1 (LYPLA1). | screening | Primary screen (LYPLA1 inhibitors in singlicate) | |
| 2233 | Counterscreen for PME1 inhibitors: fluorescence polarization-based biochemical high throughput confirmation assay for inhibitors of lysophospholipase 1 (LYPLA1). | screening | Confirmation screen (LYPLA1 inhibitors in triplicate) | |
| 2177 | Counterscreen for PME1 inhibitors: fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of lysophospholipase 2 (LYPLA2). | screening | Primary screen (LYPLA2 inhibitors in singlicate) | |
| 2232 | Counterscreen for PME1 inhibitors: fluorescence polarization-based biochemical high throughput confirmation assay to identify inhibitors of lysophospholipase 2 (LYPLA2). | screening | Confirmation screen (LYPLA2 inhibitors in triplicate) | |
| 493105 | Assay 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 enzyme | other | Confirmation screen (LYPLA1 and LYPLA2 inhibitors in singlicate) | |
| 493108 | Late stage assay provider results from the probe development effort to identify dual inhibitors of LYPLA1 and LYPLA2: fluorescence-based cell-based inhibition | other | Late stage screen (LYPLA1 and LYPLA2 inhibitors in singlicate) | |
| 493109 | Late 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 site | other | Late stage LCMS assay (LYPLA1) | |
| 493110 | Late 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 LYPLA2 | confirmatory | Late stage dose response (LYPLA1 and LYPLA2 inhibitors in triplicate) | |
| 493111 | Late 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 selectivity | other | Late stage screen (LYPLA1 and LYPLA2 inhibitors in singlicate) | |
| 493154 | Late 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 ABHD11 | confirmatory | Late stage dose response counterscreen (ABHD11 inhibitors in triplicate) | |
| 493161 | Late 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 compounds | confirmatory | Late stage dose response counterscreen (T-cell cytotoxicity in quadruplicate) | |
| 652030 | Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition and selectivity | other | ||
| 652029 | Late stage assay provider results from the probe development effort to identify selective inhibitors of LYPLA2: Fluorescence-based biochemical gel-based ABPP inhibition | other | ||
| 504892 | Late 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 ABHD11 | other | ||
| 651978 | On Hold | |||
| 651979 | On Hold | |||
| 651980 | On Hold | |||
| 651981 | On Hold | |||
| 651985 | On Hold | |||
| 651986 | On Hold | |||
| 651987 | On Hold | |||
| 651988 | On Hold | |||
| 651990 | On Hold | |||
| 651991 | On Hold | |||
| 651998 | On Hold | |||
| 652001 | On Hold | |||
| 652003 | On Hold | |||
| 652004 | On Hold | |||
| 652018 | On Hold |
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Benjamin Cravatt, TSRI
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 CA132630-01
Grant Proposal PI: Benjamin Cravatt, TSRI
External Assay ID: ABHD11_INH_LCMS_SILAC
Name: Late 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 ABHD11.
Description:
Protein palmitoylation is an essential post-translational modification necessary for trafficking and localization of regulatory proteins that play key roles in cell growth and signaling. Numerous proteins have been identified as targets of palmitoylation, including cytoskeletal proteins, kinases, receptors, and other proteins involved in various aspects of cellular signaling and homeostasis (1). Using a global chemo-proteomic method for the metabolic incorporation and identification of palmitoylated proteins, we were able to identify hundreds of palmitoylated proteins, revealing palmitoylation as a widespread post-translational modification (PTM) (2). Palmitoylation involves an acyl-thioester linkage to specific cysteines (3,4). Given the labile properties of thioesters, palmitoylation is potentially reversible and may be regulated in a manner analogous to other PTMs (e.g., phosphorylation). As such, identification of proteins responsible for the dynamic modulation of palmitoylation is paramount to understanding its patho/physiological roles. For example, multiple oncogenes, including HRAS and SRC, require palmitoylation for malignant transformation (5), suggesting protein palmitoyl thioesterases may have tumor suppressor activity required to repress aberrant growth signaling. More than a decade ago, the cytosolic serine hydrolase acyl-protein thioesterase 1 (APT1) was identified as an in vitro HRAS palmitoyl thioesterase (6). Initially classified as lysophospholipase 1 (LYPLA1) (7), the enzyme has since been demonstrated to have several hundred-fold higher activity as a protein thioesterase. While the in vitro data (6,8) provided an intriguing clue to its possible role in vivo, prior to our studies, little was known about the in vivo thioesterase activity of LYPLA1. Upon retroviral shRNA knockdown of LYPLA1, we found that HRAS was robustly hyper-palmitoylated, providing the first evidence that the endogenous enzyme is a functional protein palmitoyl thioesterase capable of regulating HRAS palmitoylation in mammalian cells. However, shRNA resulted in only an 80% reduction in LYPLA1 expression (unpublished). LYPLA2 (a.k.a. APT2) is 65% identical to LYPLA1, and also exhibits lysophospholipase activity in vitro, but its potential role as a thioesterase is unknown (9). shRNA knockdown studies of LYPLA2 revealed only partial knockdown of the enzyme, making substrate identification inconclusive (unpublished). A principle goal of post-genomic research is the determination of the molecular and cellular role of uncharacterized enzymes like LYPLA1 and LYPLA2. As such, a dual inhibitor selective for both LYPLA1 and LYPLA2 would greatly aid investigations into the biological function of these related enzymes. Several inhibitors of LYPLA1 have been described (10,11), but none of these agents have proven capable of inhibiting LYPLA1 activity in cells, and no selective inhibitors of LYPLA2 have been reported to date. To comprehensively identify LYPLA1 and LYPLA2 substrates and functionally test the role of these enzymes in dynamic de-palmitoylation and tumorigenesis, development of a high affinity inhibitor, capable of achieving temporal and more complete control over activity, is critical. Alpha/beta hydrolase domain-containing protein 11 (ABHD11) is a poorly characterized serine hydrolase; all that is known about its biology is that it is a mitochondrial enzyme (12) with broad tissue distribution, has little sequence homology to other proteins, and its gene is located in a region of chromosome 7 that is hemizygously deleted in Williams-Beuren syndrome, a rare genetic disease with symptoms that include vascular stenosis, mental retardation, and excessive sociability (13).
References:
1. Smotrys, J.E. and Linder, M.E. PALMITOYLATION OF INTRACELLULAR SIGNALING PROTEINS: Regulation and Function. Annual Review of Biochemistry, 2004. 73: 559-587.
2. Martin, B.R. and Cravatt, B.F. Large-scale profiling of protein palmitoylation in mammalian cells. Nat Methods, 2009. 6: 135-138.
3. Magee, A.I., Koyama, A.H., Malfer, C., Wen, D. and Schlesinger, M. J. Release of fatty acids from virus glycoproteins by hydroxylamine. Biochimica et Biophysica Acta (BBA) - General Subjects, 1984. 798: 156-166.
4. Rose, J.K., Adams, G.A. and Gallione, C.J. The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition. Proc Natl Acad Sci USA, 1984. 81: 2050-2054.
5. Willumsen, B.M., Cox, A.D., Solski, P.A., Der, C.J. and Buss, J.E. Novel determinants of H-Ras plasma membrane localization and transformation. Oncogene, 1996. 13: 1901-1909.
6. Duncan, J.A. and Gilman, A.G. A Cytoplasmic Acyl-Protein Thioesterase That Removes Palmitate from G Protein alpha Subunits and p21RAS. J Biol Chem, 1998. 273: 15830-15837.
7. Sugimoto, H., Hayashi, H. & Yamashita, S. Purification, cDNA cloning, and regulation of lysophospholipase from rat liver. J Biol Chem, 1996. 271: 7705-7711.
8. Hirano, T. et al. Thioesterase activity and subcellular localization of acylprotein thioesterase 1/lysophospholipase 1. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 2009. 1791: 797-805.
9. Toyoda, T., Sugimoto, H. and Yamashita, S. Sequence, expression in Escherichia coli, and characterization of lysophospholipase II. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1999. 1437: 182-193.
10. Biel, M., Deck, P., Giannis, A. and Waldmann, H. Synthesis and Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors. Chemistry - A European Journal, 2006. 12: 4121-4143.
11. Deck, P. et al. Development and Biological Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors. Angewandte Chemie International Edition, 2005. 44: 4975-4980.
12. Forner, F., et al., Quantitative proteomic comparison of rat mitochondria from muscle, heart, and liver. Mol. Cell. Proteomics, 2006. 5(4): p. 608-19.
13. Schubert, C., The genomic basis of the Williams-Beuren syndrome. Cell. Mol. Life Sci., 2009. 66(7): p. 1178-97.
Keywords:
late stage, late stage AID, assay provider, powders, LYPLA1, lysophospholipase 1, LYPLA2, lysophospholipase 2, APT1, acyl-protein thioesterase 1, APT2, acyl-protein thioesterase 2, palmitoylation, alpha/beta hydrolase domain-containing protein 11, abhydrolase domain-containing protein 11, ABHD11, oncogene, tumor suppressor, serine hydrolase, activity-based protein profiling, ABPP, gel-based ABPP, fluorophosphonate rhodamine, FP-Rh, liquid chromatography-tandem mass spectrometry, LC-MS/MS, inhibitor, in situ, cell-based assay, BW5147, murine T cells, T cells, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Benjamin Cravatt, TSRI
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R01 CA132630-01
Grant Proposal PI: Benjamin Cravatt, TSRI
External Assay ID: ABHD11_INH_LCMS_SILAC
Name: Late 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 ABHD11.
Description:
Protein palmitoylation is an essential post-translational modification necessary for trafficking and localization of regulatory proteins that play key roles in cell growth and signaling. Numerous proteins have been identified as targets of palmitoylation, including cytoskeletal proteins, kinases, receptors, and other proteins involved in various aspects of cellular signaling and homeostasis (1). Using a global chemo-proteomic method for the metabolic incorporation and identification of palmitoylated proteins, we were able to identify hundreds of palmitoylated proteins, revealing palmitoylation as a widespread post-translational modification (PTM) (2). Palmitoylation involves an acyl-thioester linkage to specific cysteines (3,4). Given the labile properties of thioesters, palmitoylation is potentially reversible and may be regulated in a manner analogous to other PTMs (e.g., phosphorylation). As such, identification of proteins responsible for the dynamic modulation of palmitoylation is paramount to understanding its patho/physiological roles. For example, multiple oncogenes, including HRAS and SRC, require palmitoylation for malignant transformation (5), suggesting protein palmitoyl thioesterases may have tumor suppressor activity required to repress aberrant growth signaling. More than a decade ago, the cytosolic serine hydrolase acyl-protein thioesterase 1 (APT1) was identified as an in vitro HRAS palmitoyl thioesterase (6). Initially classified as lysophospholipase 1 (LYPLA1) (7), the enzyme has since been demonstrated to have several hundred-fold higher activity as a protein thioesterase. While the in vitro data (6,8) provided an intriguing clue to its possible role in vivo, prior to our studies, little was known about the in vivo thioesterase activity of LYPLA1. Upon retroviral shRNA knockdown of LYPLA1, we found that HRAS was robustly hyper-palmitoylated, providing the first evidence that the endogenous enzyme is a functional protein palmitoyl thioesterase capable of regulating HRAS palmitoylation in mammalian cells. However, shRNA resulted in only an 80% reduction in LYPLA1 expression (unpublished). LYPLA2 (a.k.a. APT2) is 65% identical to LYPLA1, and also exhibits lysophospholipase activity in vitro, but its potential role as a thioesterase is unknown (9). shRNA knockdown studies of LYPLA2 revealed only partial knockdown of the enzyme, making substrate identification inconclusive (unpublished). A principle goal of post-genomic research is the determination of the molecular and cellular role of uncharacterized enzymes like LYPLA1 and LYPLA2. As such, a dual inhibitor selective for both LYPLA1 and LYPLA2 would greatly aid investigations into the biological function of these related enzymes. Several inhibitors of LYPLA1 have been described (10,11), but none of these agents have proven capable of inhibiting LYPLA1 activity in cells, and no selective inhibitors of LYPLA2 have been reported to date. To comprehensively identify LYPLA1 and LYPLA2 substrates and functionally test the role of these enzymes in dynamic de-palmitoylation and tumorigenesis, development of a high affinity inhibitor, capable of achieving temporal and more complete control over activity, is critical. Alpha/beta hydrolase domain-containing protein 11 (ABHD11) is a poorly characterized serine hydrolase; all that is known about its biology is that it is a mitochondrial enzyme (12) with broad tissue distribution, has little sequence homology to other proteins, and its gene is located in a region of chromosome 7 that is hemizygously deleted in Williams-Beuren syndrome, a rare genetic disease with symptoms that include vascular stenosis, mental retardation, and excessive sociability (13).
References:
1. Smotrys, J.E. and Linder, M.E. PALMITOYLATION OF INTRACELLULAR SIGNALING PROTEINS: Regulation and Function. Annual Review of Biochemistry, 2004. 73: 559-587.
2. Martin, B.R. and Cravatt, B.F. Large-scale profiling of protein palmitoylation in mammalian cells. Nat Methods, 2009. 6: 135-138.
3. Magee, A.I., Koyama, A.H., Malfer, C., Wen, D. and Schlesinger, M. J. Release of fatty acids from virus glycoproteins by hydroxylamine. Biochimica et Biophysica Acta (BBA) - General Subjects, 1984. 798: 156-166.
4. Rose, J.K., Adams, G.A. and Gallione, C.J. The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition. Proc Natl Acad Sci USA, 1984. 81: 2050-2054.
5. Willumsen, B.M., Cox, A.D., Solski, P.A., Der, C.J. and Buss, J.E. Novel determinants of H-Ras plasma membrane localization and transformation. Oncogene, 1996. 13: 1901-1909.
6. Duncan, J.A. and Gilman, A.G. A Cytoplasmic Acyl-Protein Thioesterase That Removes Palmitate from G Protein alpha Subunits and p21RAS. J Biol Chem, 1998. 273: 15830-15837.
7. Sugimoto, H., Hayashi, H. & Yamashita, S. Purification, cDNA cloning, and regulation of lysophospholipase from rat liver. J Biol Chem, 1996. 271: 7705-7711.
8. Hirano, T. et al. Thioesterase activity and subcellular localization of acylprotein thioesterase 1/lysophospholipase 1. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 2009. 1791: 797-805.
9. Toyoda, T., Sugimoto, H. and Yamashita, S. Sequence, expression in Escherichia coli, and characterization of lysophospholipase II. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1999. 1437: 182-193.
10. Biel, M., Deck, P., Giannis, A. and Waldmann, H. Synthesis and Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors. Chemistry - A European Journal, 2006. 12: 4121-4143.
11. Deck, P. et al. Development and Biological Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors. Angewandte Chemie International Edition, 2005. 44: 4975-4980.
12. Forner, F., et al., Quantitative proteomic comparison of rat mitochondria from muscle, heart, and liver. Mol. Cell. Proteomics, 2006. 5(4): p. 608-19.
13. Schubert, C., The genomic basis of the Williams-Beuren syndrome. Cell. Mol. Life Sci., 2009. 66(7): p. 1178-97.
Keywords:
late stage, late stage AID, assay provider, powders, LYPLA1, lysophospholipase 1, LYPLA2, lysophospholipase 2, APT1, acyl-protein thioesterase 1, APT2, acyl-protein thioesterase 2, palmitoylation, alpha/beta hydrolase domain-containing protein 11, abhydrolase domain-containing protein 11, ABHD11, oncogene, tumor suppressor, serine hydrolase, activity-based protein profiling, ABPP, gel-based ABPP, fluorophosphonate rhodamine, FP-Rh, liquid chromatography-tandem mass spectrometry, LC-MS/MS, inhibitor, in situ, cell-based assay, BW5147, murine T cells, T cells, 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 | ABHD11 | 1 | Abhydrolase domain containing 11 [Mus musculus] [gi:47682716] | Taxonomy id: 10090 Gene id: 68758 | |||
| 2 | ABHD6 | 1 | Abhydrolase domain containing 6 [Mus musculus] [gi:20073260] | Taxonomy id: 10090 Gene id: 66082 | |||
| 3 | ACOT1 | 1 | Acyl-CoA thioesterase 1 [Mus musculus] [gi:20380528] | Taxonomy id: 10090 Gene id: 26897 | |||
| 4 | DPP4 | 1 | dipeptidylpeptidase 4 [Mus musculus] [gi:123233284] | Taxonomy id: 10090 Gene id: 13482 | |||
| 5 | ACOT2 | 1 | Acyl-CoA thioesterase 2 [Mus musculus] [gi:39992610] | Taxonomy id: 10090 Gene id: 171210 | |||
| 6 | PREPL | 1 | Prepl protein [Mus musculus] [gi:13435484] | Taxonomy id: 10090 Gene id: 213760 | |||
| 7 | FAM108B1 | 1 | Fam108b protein [Mus musculus] [gi:61403362] | Taxonomy id: 10090 Gene id: 226016 | |||
| 8 | PAFAH2 | 1 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] [gi:225579137] | Taxonomy id: 10090 Gene id: 100163 | |||
| 9 | CTSA | 1 | cathepsin A [Mus musculus] [gi:123240058] | Taxonomy id: 10090 Gene id: 19025 | |||
| 10 | ABHD12 | 1 | abhydrolase domain containing 12 [Mus musculus] [gi:123241061] | Taxonomy id: 10090 Gene id: 76192 | |||
| 11 | PNPLA8 | 1 | Patatin-like phospholipase domain containing 8 [Mus musculus] [gi:117558667] | Taxonomy id: 10090 Gene id: 67452 | |||
| 12 | AADACL1 | 1 | Arylacetamide deacetylase-like 1 [Mus musculus] [gi:52139053] | Taxonomy id: 10090 Gene id: 320024 | |||
| 13 | DPP7 | 1 | dipeptidylpeptidase 7 [Mus musculus] [gi:123227495] | Taxonomy id: 10090 Gene id: 83768 | |||
| 14 | PAFAH1B3 | 1 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] [gi:6679201] | Taxonomy id: 10090 Gene id: 18476 | |||
| 15 | ABHD10 | 1 | Abhydrolase domain containing 10 [Mus musculus] [gi:37231537] | Taxonomy id: 10090 Gene id: 213012 | |||
| 16 | ESD | 1 | Esterase D/formylglutathione hydrolase [Mus musculus] [gi:55777188] | Taxonomy id: 10090 Gene id: 13885 | |||
| 17 | IAH1 | 1 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] [gi:27754071] | Taxonomy id: 10090 Gene id: 67732 | |||
| 18 | APEH | 1 | Apeh protein [Mus musculus] [gi:20072022] | Taxonomy id: 10090 Gene id: 235606 | |||
| 19 | LIPA | 1 | lysosomal acid lipase 1 [Mus musculus] [gi:148709804] | Taxonomy id: 10090 Gene id: 16889 | |||
| 20 | PNPLA6 | 1 | Pnpla6 protein [Mus musculus] [gi:34784201] | Taxonomy id: 10090 Gene id: 50767 | |||
| 21 | PLA2G6 | 1 | Pla2g6 protein [Mus musculus] [gi:34784362] | Taxonomy id: 10090 Gene id: 53357 | |||
| 22 | PREP | 1 | prolyl endopeptidase [Mus musculus] [gi:148673089] | Taxonomy id: 10090 Gene id: 19072 | |||
| 23 | PRCP | 1 | Prcp protein [Mus musculus] [gi:32967631] | Taxonomy id: 10090 Gene id: 72461 | |||
| 24 | PPME1 | 1 | protein phosphatase methylesterase 1 [Mus musculus] [gi:30794138] | Taxonomy id: 10090 Gene id: 72590 | |||
| 25 | ACOT7 | 1 | acyl-CoA thioesterase 7 [Mus musculus] [gi:123857891] | Taxonomy id: 10090 Gene id: 70025 | |||
| 26 | LACTB | 1 | Lactamase, beta [Mus musculus] [gi:187955464] | Taxonomy id: 10090 Gene id: 80907 | |||
| 27 | LYPLA3 | 1 | lysophospholipase 3, isoform CRA_a [Mus musculus] [gi:148679401] | Taxonomy id: 10090 Gene id: 192654 | |||
| 28 | SERHL | 1 | Serine hydrolase-like [Mus musculus] [gi:127797596] | Taxonomy id: 10090 Gene id: 68607 | |||
| 29 | PARL | 1 | presenilin associated, rhomboid-like [Mus musculus] [gi:148665146] | Taxonomy id: 10090 Gene id: 381038 | |||
| 30 | DPP8 | 1 | Dipeptidylpeptidase 8 [Mus musculus] [gi:37590654] | Taxonomy id: 10090 Gene id: 74388 | |||
| 31 | BAT5 | 1 | abhydrolase domain-containing protein 16A [Mus musculus] [gi:30519896] | Taxonomy id: 10090 Gene id: 193742 | |||
| 32 | LYPLA1 | 1 | Lypla1 [Mus musculus] [gi:71059731] | Taxonomy id: 10090 Gene id: 18777 | |||
| 33 | SIAE | 1 | sialic acid acetylesterase [Mus musculus] [gi:148693491] | Taxonomy id: 10090 Gene id: 22619 | |||
| 34 | FAAH | 1 | fatty acid amide hydrolase [Mus musculus] [gi:123253900] | Taxonomy id: 10090 Gene id: 14073 | |||
| 35 | DPP9 | 1 | dipeptidyl peptidase 9 [Mus musculus] [gi:255003757] | Taxonomy id: 10090 Gene id: 224897 | |||
| 36 | FASN | 1 | fatty acid synthase [Mus musculus] [gi:123288587] | Taxonomy id: 10090 Gene id: 14104 | |||
| 37 | LYPLA2 | 1 | lysophospholipase 2 [Mus musculus] [gi:123122209] | Taxonomy id: 10090 Gene id: 26394 | |||
| 38 | PAFAH1B2 | 1 | platelet-activating factor acetylhydrolase isoform Ib beta subunit [Mus musculus] [gi:1373363] | Taxonomy id: 10090 Gene id: 18475 | |||
| 39 | DDHD1 | 1 | Ddhd1 protein [Mus musculus] [gi:27694042] | Taxonomy id: 10090 Gene id: 114874 | |||
§ Panel component ID.
Protocol
Assay Overview:
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using stable isotope labeling with amino acids in cell culture (SILAC) ABPP. In this assay, cultured BW5147-derived murine T-cells are metabolically labeled with light or heavy amino acids. Light and heavy cells are treated with inhibitor and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with FP-biotin, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by 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 (light) sample relative to the DMSO-treated (heavy) sample, giving a smaller light/heavy ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Stable isotope labeling with amino acids in cell culture (SILAC)
BW5147-derived murine T-cell hybridoma cells were initially grown for 6 passages in either light or heavy SILAC RPMI 1640 media supplemented with 10% dialyzed FCS and 1x PenStrep Glutamine. Light media was supplemented with 100 ug/mL L-arginine (Sigma) and 100 ug/mL L-lysine (Sigma). Heavy media was supplemented with 100 ug/mL [13C615N4]-L-Arginine (Isotek) and 100 ug/mL [13C615N2]-L-Lysine (Isotek). Cells were treated with 3 nM test compound (5 uL of a 1000x stock in DMSO) for 4 hours at 37 C. Cells were harvested, washed 4 times with 10 mL DPBS, and homogenized by sonication in DPBS. The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration was adjusted to 1 mg/mL with DPBS.
Sample preparation for ABPP-SILAC
The light and heavy proteomes were labeled with 7 uM of FP-biotin (500 uL total reaction volume) for 1.5 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. The proteomes were desalted over PD-10 desalting columns (GE Healthcare) and FP-labeled proteins were enriched with streptavidin beads. The beads were washed with 1% SDS in PBS (1x), PBS (3x), and H2O (3x), then resuspended in 6 M urea, reduced with DTT for 15 minutes at 60 C, and alkylated with iodoacetamide for 30 minutes at 25 C in the dark. On-bead digestions were performed for 12 hours at 37 C with trypsin (Promega; 4 uL of 0.5 ug/uL) in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% formic acid, pressure-loaded on to a biphasic (strong cation exchange/reverse phase) capillary column and analyzed as described below.
LC-MS/MS analysis
Digested and acidified peptide mixtures were analyzed by two-dimensional liquid chromatography (2D-LC) separation in combination with tandem mass spectrometry using an Agilent 1100-series quaternary pump and Thermo Scientific LTQ Orbitrap 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 (60 s, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 7 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.1; publicly available at http://fields.scripps.edu/?q=content/download). MS2 spectra data were searched using the SEQUEST algorithm (Version 3.0) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. SEQUEST searches allowed for variable oxidation of methionine (+16), static modification of cysteine residues (+57 due to alkylation), and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0.41) using the --trypstat option, which applies different statistical models for the analysis of tryptic, half-tryptic, non-tryptic peptides. DTASelect 2.0 uses a quadratic discriminant analysis to achieve a user-defined maximum peptide false positive rate; the default parameters (maximum false positive rate of 2%) was used for the search; however, the actual false positive rate was much lower (1%). Ratios of Light/Heavy peaks were calculated using in-house software; reported ratios represent the mean of all unique, quantified peptides per protein.
Ratio = Average( AUC_light / AUC_heavy ) calculated for all unique peptides
Where:
AUC_light is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUC_heavy 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 ABHD11 and inactive for all anti-target serine hydrolases tested.
Active compounds were given a score of 100 and inactive compounds were given a score of 0.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
BW5147-derived murine T-cells (provided by Assay Provider)
SILAC RPMI 1640 media (Thermo 89984)
dialyzed FCS (Gemini 100-108)
1x PenStrep Glutamine (CellGro 30-002-CI)
L-Arginine (Sigma A6969)
L-Lysine (Sigma L9037)
[13C615N4]-L-Arginine (Sigma 608033)
[13C615N2]-L-Lysine (Sigma 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
PD-10 desalting columns (GE Healthcare 17-0851-01)
SDS (Sigma L6026)
Urea (Fisher U15-3)
DTT (Sigma 43815)
Iodoacetamide(Sigma I1149)
Trypsin (Promega V5111)
CaCl2 (Sigma C1016)
streptavidin beads (Pierce 20349)
Fused-silica (Agilent 160-2635-10)
Aqua C18 (Phenomenex 04A-4299)
Acetonitrile (Fisher A955-4)
Millipore-filtered Water
Formic acid (Fluka 06440)
Triton-X100 (Fisher AC21568-0010)
The purpose of this assay is to determine the selectivity profile of powder samples of test compounds using stable isotope labeling with amino acids in cell culture (SILAC) ABPP. In this assay, cultured BW5147-derived murine T-cells are metabolically labeled with light or heavy amino acids. Light and heavy cells are treated with inhibitor and DMSO, respectively, in situ. Cells are lysed, proteomes are treated with FP-biotin, and combined in a 1:1 (w/w) ratio. Biotinylated proteins are enriched, trypsinized, and analyzed by 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 (light) sample relative to the DMSO-treated (heavy) sample, giving a smaller light/heavy ratio for each protein. Proteins not targeted by inhibitors would be expected to have a ratio of 1.
Protocol Summary:
Stable isotope labeling with amino acids in cell culture (SILAC)
BW5147-derived murine T-cell hybridoma cells were initially grown for 6 passages in either light or heavy SILAC RPMI 1640 media supplemented with 10% dialyzed FCS and 1x PenStrep Glutamine. Light media was supplemented with 100 ug/mL L-arginine (Sigma) and 100 ug/mL L-lysine (Sigma). Heavy media was supplemented with 100 ug/mL [13C615N4]-L-Arginine (Isotek) and 100 ug/mL [13C615N2]-L-Lysine (Isotek). Cells were treated with 3 nM test compound (5 uL of a 1000x stock in DMSO) for 4 hours at 37 C. Cells were harvested, washed 4 times with 10 mL DPBS, and homogenized by sonication in DPBS. The soluble and membrane fractions were isolated by centrifugation (100K x g, 45 minutes) and the protein concentration was adjusted to 1 mg/mL with DPBS.
Sample preparation for ABPP-SILAC
The light and heavy proteomes were labeled with 7 uM of FP-biotin (500 uL total reaction volume) for 1.5 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. The proteomes were desalted over PD-10 desalting columns (GE Healthcare) and FP-labeled proteins were enriched with streptavidin beads. The beads were washed with 1% SDS in PBS (1x), PBS (3x), and H2O (3x), then resuspended in 6 M urea, reduced with DTT for 15 minutes at 60 C, and alkylated with iodoacetamide for 30 minutes at 25 C in the dark. On-bead digestions were performed for 12 hours at 37 C with trypsin (Promega; 4 uL of 0.5 ug/uL) in the presence of 2 mM CaCl2. Peptide samples were acidified to a final concentration of 5% formic acid, pressure-loaded on to a biphasic (strong cation exchange/reverse phase) capillary column and analyzed as described below.
LC-MS/MS analysis
Digested and acidified peptide mixtures were analyzed by two-dimensional liquid chromatography (2D-LC) separation in combination with tandem mass spectrometry using an Agilent 1100-series quaternary pump and Thermo Scientific LTQ Orbitrap 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 (60 s, repeat of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) was followed by 7 MS2 scans of the most abundant ions. The MS2 spectra data were extracted from the raw file using RAW Xtractor (version 1.9.1; publicly available at http://fields.scripps.edu/?q=content/download). MS2 spectra data were searched using the SEQUEST algorithm (Version 3.0) against the latest version of the mouse IPI database concatenated with the reversed database for assessment of false-discovery rates. SEQUEST searches allowed for variable oxidation of methionine (+16), static modification of cysteine residues (+57 due to alkylation), and no enzyme specificity. The resulting MS2 spectra matches were assembled into protein identifications and filtered using DTASelect (version 2.0.41) using the --trypstat option, which applies different statistical models for the analysis of tryptic, half-tryptic, non-tryptic peptides. DTASelect 2.0 uses a quadratic discriminant analysis to achieve a user-defined maximum peptide false positive rate; the default parameters (maximum false positive rate of 2%) was used for the search; however, the actual false positive rate was much lower (1%). Ratios of Light/Heavy peaks were calculated using in-house software; reported ratios represent the mean of all unique, quantified peptides per protein.
Ratio = Average( AUC_light / AUC_heavy ) calculated for all unique peptides
Where:
AUC_light is the area-under-the-curve for the light peptide pair from cells treated with test compound.
AUC_heavy 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 ABHD11 and inactive for all anti-target serine hydrolases tested.
Active compounds were given a score of 100 and inactive compounds were given a score of 0.
The PubChem Activity Score range for active compounds is 100-100. There are no inactive compounds.
List of Reagents:
BW5147-derived murine T-cells (provided by Assay Provider)
SILAC RPMI 1640 media (Thermo 89984)
dialyzed FCS (Gemini 100-108)
1x PenStrep Glutamine (CellGro 30-002-CI)
L-Arginine (Sigma A6969)
L-Lysine (Sigma L9037)
[13C615N4]-L-Arginine (Sigma 608033)
[13C615N2]-L-Lysine (Sigma 608041)
DPBS (Cellgro 20-031-CV)
FP-biotin (provided by Assay Provider)
PD-10 desalting columns (GE Healthcare 17-0851-01)
SDS (Sigma L6026)
Urea (Fisher U15-3)
DTT (Sigma 43815)
Iodoacetamide(Sigma I1149)
Trypsin (Promega V5111)
CaCl2 (Sigma C1016)
streptavidin beads (Pierce 20349)
Fused-silica (Agilent 160-2635-10)
Aqua C18 (Phenomenex 04A-4299)
Acetonitrile (Fisher A955-4)
Millipore-filtered Water
Formic acid (Fluka 06440)
Triton-X100 (Fisher AC21568-0010)
Comment
This assay was performed by the assay provider with powder samples of compounds.
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 [ABHD11] | One of Active or Inactive | 1 | Abhydrolase domain containing 11 [Mus musculus] | | Outcome | ||
| 2 | Average Silac Ratio [ABHD11] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 1 | | Float | ratio | ||
| 3 | Standard Error [ABHD11] | Calculated standard error for mean | 1 | | Float | |||
| 4 | Outcome [ABHD6] | One of Active or Inactive | 2 | Abhydrolase domain containing 6 [Mus musculus] | | Outcome | ||
| 5 | Average Silac Ratio [ABHD6] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 2 | | Float | ratio | ||
| 6 | Standard Error [ABHD6] | Calculated standard error for mean | 2 | | Float | |||
| 7 | Outcome [ACOT1] | One of Active or Inactive | 3 | Acyl-CoA thioesterase 1 [Mus musculus] | | Outcome | ||
| 8 | Average Silac Ratio [ACOT1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 3 | | Float | ratio | ||
| 9 | Standard Error [ACOT1] | Calculated standard error for mean | 3 | | Float | |||
| 10 | Outcome [DPP4] | One of Active or Inactive | 4 | dipeptidylpeptidase 4 [Mus musculus] | | Outcome | ||
| 11 | Average Silac Ratio [DPP4] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 4 | | Float | ratio | ||
| 12 | Standard Error [DPP4] | Calculated standard error for mean | 4 | | Float | |||
| 13 | Outcome [ACOT2] | One of Active or Inactive | 5 | Acyl-CoA thioesterase 2 [Mus musculus] | | Outcome | ||
| 14 | Average Silac Ratio [ACOT2] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 5 | | Float | ratio | ||
| 15 | Standard Error [ACOT2] | Calculated standard error for mean | 5 | | Float | |||
| 16 | Outcome [PREPL] | One of Active or Inactive | 6 | Prepl protein [Mus musculus] | | Outcome | ||
| 17 | Average Silac Ratio [PREPL] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 6 | | Float | ratio | ||
| 18 | Standard Error [PREPL] | Calculated standard error for mean | 6 | | Float | |||
| 19 | Outcome [FAM108B1] | One of Active or Inactive | 7 | Fam108b protein [Mus musculus] | | Outcome | ||
| 20 | Average Silac Ratio [FAM108B1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 7 | | Float | ratio | ||
| 21 | Standard Error [FAM108B1] | Calculated standard error for mean | 7 | | Float | |||
| 22 | Outcome [PAFAH2] | One of Active or Inactive | 8 | platelet-activating factor acetylhydrolase 2, cytoplasmic [Mus musculus] | | Outcome | ||
| 23 | Average Silac Ratio [PAFAH2] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 8 | | Float | ratio | ||
| 24 | Standard Error [PAFAH2] | Calculated standard error for mean | 8 | | Float | |||
| 25 | Outcome [CTSA] | One of Active or Inactive | 9 | cathepsin A [Mus musculus] | | Outcome | ||
| 26 | Average Silac Ratio [CTSA] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 9 | | Float | ratio | ||
| 27 | Standard Error [CTSA] | Calculated standard error for mean | 9 | | Float | |||
| 28 | Outcome [ABHD12] | One of Active or Inactive | 10 | abhydrolase domain containing 12 [Mus musculus] | | Outcome | ||
| 29 | Average Silac Ratio [ABHD12] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 10 | | Float | ratio | ||
| 30 | Standard Error [ABHD12] | Calculated standard error for mean | 10 | | Float | |||
| 31 | Outcome [PNPLA8] | One of Active or Inactive | 11 | Patatin-like phospholipase domain containing 8 [Mus musculus] | | Outcome | ||
| 32 | Average Silac Ratio [PNPLA8] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 11 | | Float | ratio | ||
| 33 | Standard Error [PNPLA8] | Calculated standard error for mean | 11 | | Float | |||
| 34 | Outcome [AADACL1] | One of Active or Inactive | 12 | Arylacetamide deacetylase-like 1 [Mus musculus] | | Outcome | ||
| 35 | Average Silac Ratio [AADACL1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 12 | | Float | ratio | ||
| 36 | Standard Error [AADACL1] | Calculated standard error for mean | 12 | | Float | |||
| 37 | Outcome [DPP7] | One of Active or Inactive | 13 | dipeptidylpeptidase 7 [Mus musculus] | | Outcome | ||
| 38 | Average Silac Ratio [DPP7] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 13 | | Float | ratio | ||
| 39 | Standard Error [DPP7] | Calculated standard error for mean | 13 | | Float | |||
| 40 | Outcome [PAFAH1B3] | One of Active or Inactive | 14 | platelet-activating factor acetylhydrolase IB subunit gamma [Mus musculus] | | Outcome | ||
| 41 | Average Silac Ratio [PAFAH1B3] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 14 | | Float | ratio | ||
| 42 | Standard Error [PAFAH1B3] | Calculated standard error for mean | 14 | | Float | |||
| 43 | Outcome [ABHD10] | One of Active or Inactive | 15 | Abhydrolase domain containing 10 [Mus musculus] | | Outcome | ||
| 44 | Average Silac Ratio [ABHD10] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 15 | | Float | ratio | ||
| 45 | Standard Error [ABHD10] | Calculated standard error for mean | 15 | | Float | |||
| 46 | Outcome [ESD] | One of Active or Inactive | 16 | Esterase D/formylglutathione hydrolase [Mus musculus] | | Outcome | ||
| 47 | Average Silac Ratio [ESD] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 16 | | Float | ratio | ||
| 48 | Standard Error [ESD] | Calculated standard error for mean | 16 | | Float | |||
| 49 | Outcome [IAH1] | One of Active or Inactive | 17 | isoamyl acetate-hydrolyzing esterase 1 homolog [Mus musculus] | | Outcome | ||
| 50 | Average Silac Ratio [IAH1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 17 | | Float | ratio | ||
| 51 | Standard Error [IAH1] | Calculated standard error for mean | 17 | | Float | |||
| 52 | Outcome [APEH] | One of Active or Inactive | 18 | Apeh protein [Mus musculus] | | Outcome | ||
| 53 | Average Silac Ratio [APEH] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 18 | | Float | ratio | ||
| 54 | Standard Error [APEH] | Calculated standard error for mean | 18 | | Float | |||
| 55 | Outcome [LIPA] | One of Active or Inactive | 19 | lysosomal acid lipase 1 [Mus musculus] | | Outcome | ||
| 56 | Average Silac Ratio [LIPA] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 19 | | Float | ratio | ||
| 57 | Standard Error [LIPA] | Calculated standard error for mean | 19 | | Float | |||
| 58 | Outcome [PNPLA6] | One of Active or Inactive | 20 | Pnpla6 protein [Mus musculus] | | Outcome | ||
| 59 | Average Silac Ratio [PNPLA6] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 20 | | Float | ratio | ||
| 60 | Standard Error [PNPLA6] | Calculated standard error for mean | 20 | | Float | |||
| 61 | Outcome [PLA2G6] | One of Active or Inactive | 21 | Pla2g6 protein [Mus musculus] | | Outcome | ||
| 62 | Average Silac Ratio [PLA2G6] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 21 | | Float | ratio | ||
| 63 | Standard Error [PLA2G6] | Calculated standard error for mean | 21 | | Float | |||
| 64 | Outcome [PREP] | One of Active or Inactive | 22 | prolyl endopeptidase [Mus musculus] | | Outcome | ||
| 65 | Average Silac Ratio [PREP] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 22 | | Float | ratio | ||
| 66 | Standard Error [PREP] | Calculated standard error for mean | 22 | | Float | |||
| 67 | Outcome [PRCP] | One of Active or Inactive | 23 | Prcp protein [Mus musculus] | | Outcome | ||
| 68 | Average Silac Ratio [PRCP] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 23 | | Float | ratio | ||
| 69 | Standard Error [PRCP] | Calculated standard error for mean | 23 | | Float | |||
| 70 | Outcome [PPME1] | One of Active or Inactive | 24 | protein phosphatase methylesterase 1 [Mus musculus] | | Outcome | ||
| 71 | Average Silac Ratio [PPME1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 24 | | Float | ratio | ||
| 72 | Standard Error [PPME1] | Calculated standard error for mean | 24 | | Float | |||
| 73 | Outcome [ACOT7] | One of Active or Inactive | 25 | acyl-CoA thioesterase 7 [Mus musculus] | | Outcome | ||
| 74 | Average Silac Ratio [ACOT7] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 25 | | Float | ratio | ||
| 75 | Standard Error [ACOT7] | Calculated standard error for mean | 25 | | Float | |||
| 76 | Outcome [LACTB] | One of Active or Inactive | 26 | Lactamase, beta [Mus musculus] | | Outcome | ||
| 77 | Average Silac Ratio [LACTB] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 26 | | Float | ratio | ||
| 78 | Standard Error [LACTB] | Calculated standard error for mean | 26 | | Float | |||
| 79 | Outcome [LYPLA3] | One of Active or Inactive | 27 | lysophospholipase 3, isoform CRA_a [Mus musculus] | | Outcome | ||
| 80 | Average Silac Ratio [LYPLA3] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 27 | | Float | ratio | ||
| 81 | Standard Error [LYPLA3] | Calculated standard error for mean | 27 | | Float | |||
| 82 | Outcome [SERHL] | One of Active or Inactive | 28 | Serine hydrolase-like [Mus musculus] | | Outcome | ||
| 83 | Average Silac Ratio [SERHL] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 28 | | Float | ratio | ||
| 84 | Standard Error [SERHL] | Calculated standard error for mean | 28 | | Float | |||
| 85 | Outcome [PARL] | One of Active or Inactive | 29 | presenilin associated, rhomboid-like [Mus musculus] | | Outcome | ||
| 86 | Average Silac Ratio [PARL] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 29 | | Float | ratio | ||
| 87 | Standard Error [PARL] | Calculated standard error for mean | 29 | | Float | |||
| 88 | Outcome [DPP8] | One of Active or Inactive | 30 | Dipeptidylpeptidase 8 [Mus musculus] | | Outcome | ||
| 89 | Average Silac Ratio [DPP8] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 30 | | Float | ratio | ||
| 90 | Standard Error [DPP8] | Calculated standard error for mean | 30 | | Float | |||
| 91 | Outcome [BAT5] | One of Active or Inactive | 31 | abhydrolase domain-containing protein 16A [Mus musculus] | | Outcome | ||
| 92 | Average Silac Ratio [BAT5] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 31 | | Float | ratio | ||
| 93 | Standard Error [BAT5] | Calculated standard error for mean | 31 | | Float | |||
| 94 | Outcome [LYPLA1] | One of Active or Inactive | 32 | Lypla1 [Mus musculus] | | Outcome | ||
| 95 | Average Silac Ratio [LYPLA1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 32 | | Float | ratio | ||
| 96 | Standard Error [LYPLA1] | Calculated standard error for mean | 32 | | Float | |||
| 97 | Outcome [SIAE] | One of Active or Inactive | 33 | sialic acid acetylesterase [Mus musculus] | | Outcome | ||
| 98 | Average Silac Ratio [SIAE] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 33 | | Float | ratio | ||
| 99 | Standard Error [SIAE] | Calculated standard error for mean | 33 | | Float | |||
| 100 | Outcome [FAAH] | One of Active or Inactive | 34 | fatty acid amide hydrolase [Mus musculus] | | Outcome | ||
| 101 | Average Silac Ratio [FAAH] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 34 | | Float | ratio | ||
| 102 | Standard Error [FAAH] | Calculated standard error for mean | 34 | | Float | |||
| 103 | Outcome [DPP9] | One of Active or Inactive | 35 | dipeptidyl peptidase 9 [Mus musculus] | | Outcome | ||
| 104 | Average Silac Ratio [DPP9] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 35 | | Float | ratio | ||
| 105 | Standard Error [DPP9] | Calculated standard error for mean | 35 | | Float | |||
| 106 | Outcome [FASN] | One of Active or Inactive | 36 | fatty acid synthase [Mus musculus] | | Outcome | ||
| 107 | Average Silac Ratio [FASN] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 36 | | Float | ratio | ||
| 108 | Standard Error [FASN] | Calculated standard error for mean | 36 | | Float | |||
| 109 | Outcome [LYPLA2] | One of Active or Inactive | 37 | lysophospholipase 2 [Mus musculus] | | Outcome | ||
| 110 | Average Silac Ratio [LYPLA2] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 37 | | Float | ratio | ||
| 111 | Standard Error [LYPLA2] | Calculated standard error for mean | 37 | | Float | |||
| 112 | Outcome [PAFAH1B2] | One of Active or Inactive | 38 | platelet-activating factor acetylhydrolase isoform Ib beta subunit [Mus musculus] | | Outcome | ||
| 113 | Average Silac Ratio [PAFAH1B2] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 38 | | Float | ratio | ||
| 114 | Standard Error [PAFAH1B2] | Calculated standard error for mean | 38 | | Float | |||
| 115 | Outcome [DDHD1] | One of Active or Inactive | 39 | Ddhd1 protein [Mus musculus] | | Outcome | ||
| 116 | Average Silac Ratio [DDHD1] | Calculated light/heavy (inhibitor-treated/DMSO) ratio for each protein as mean for all unique peptides quantified | 39 | | Float | ratio | ||
| 117 | Standard Error [DDHD1] | Calculated standard error for mean | 39 | | Float |
Additional Information
Grant Number: 1 R01 CA132630-01
Classification
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