AADAC is a single-pass type II endoplasmic-reticulum membrane serine hydrolase/deacetylase expressed most strongly in liver and gastrointestinal tissues. Its lumen-facing catalytic region hydrolyzes small acyl groups from structurally diverse amide and ester xenobiotics, thereby altering drug activation, exposure, and toxicity in a substrate-dependent manner. AADAC also contributes to ER-associated neutral-lipid mobilization: ortholog experiments directly support hydrolysis of endogenous diacylglycerol, whereas human-cell perturbations establish effects on lipid storage and lipoprotein production without yet defining the direct human lipid substrate. Cholesteryl-acetate and pineal-indole hydrolysis are experimentally supported context-specific activities whose physiological importance in humans remains unresolved.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0046340 diacylglycerol catabolic process | IBA GO_REF:0000033 | ACCEPT | Summary: AADAC-family evidence supports hydrolysis of endogenously synthesized diacylglycerol and therefore a direct role in diacylglycerol catabolism. The exact biochemical evidence is strongest for mouse Aadac expressed in rat hepatoma cells; human AADAC has complementary cellular evidence for hepatic neutral-lipid lipolysis. Reason: The mouse ortholog is an appropriate close donor and was reported to hydrolyze endogenously synthesized diacylglycerol (PMID:19654421). Human AADAC knockdown and re-expression alter cellular triacylglycerol lipolysis (PMID:23542347), supporting conservation while leaving purified-human substrate kinetics as an evidence gap. Propagation Review Root cause: NO FAILURE CORE Sources checked: MGI:MGI:1915008 · mouse Aadac SUPPORTS TRANSFER Close ortholog with experimental diacylglycerol-hydrolysis evidence. PANTHER:PTN002745055 · AADAC family tree node SUPPORTS TRANSFER Phylogenetic node connecting the experimentally supported mouse and human proteins. Supporting Evidence: PMID:19654421 Although AADA shares similarity with HSL and like HSL showed hydrolytic activity toward endogenously synthesized DG, we did not observe any in vitro activity of AADA toward membrane-associated TG or CE. PMID:23542347 The re-introduction of AADAC to infected cells restored cellular TG lipolysis, indicating a role for HCV-mediated downregulation of AADAC in this process. |
| GO:0120516 diacylglycerol lipase activity | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetic inference of diacylglycerol lipase activity is consistent with experimental hydrolysis of endogenously synthesized diacylglycerol by mouse Aadac and AADAC-dependent neutral-lipid lipolysis in human hepatoma cells. Reason: The term captures the best-resolved endogenous lipid substrate reported for the ortholog. Evidence for the exact reaction is direct in the mouse-ortholog cell system (PMID:19654421), whereas the human data are cellular rather than a purified human enzyme assay (PMID:23542347); that boundary is retained rather than treated as evidence against a conservative IBA transfer. Propagation Review Root cause: NO FAILURE CORE Sources checked: MGI:MGI:1915008 · mouse Aadac SUPPORTS TRANSFER Close ortholog with experimental endogenous diacylglycerol-hydrolysis evidence. PANTHER:PTN002745055 · AADAC family tree node SUPPORTS TRANSFER The family placement supports transfer to human AADAC. Supporting Evidence: PMID:19654421 Based on our findings, we propose a model where AADA hydrolyzes DG synthesized within the ER compartment. PMID:23542347 Defective lipolysis of cellular TG stores and VLDL production were also observed in HuH7.5 cells stably expressing a short hairpin RNA targeting AADAC expression, proving AADAC deficiency contributes to these defective pathways. |
| GO:0005789 endoplasmic reticulum membrane | IBA GO_REF:0000033 | ACCEPT | Summary: AADAC is a single-pass type II endoplasmic-reticulum membrane enzyme whose catalytic domain faces the lumen. The IBA localization agrees with independent human microsomal and endoplasmic-reticulum evidence. Reason: Human recombinant and tissue-fractionation work places AADAC in the endoplasmic reticulum and finds activity in liver microsomes rather than cytosol (PMID:19339378). The mouse ortholog therefore supports, rather than drives, an otherwise well-established human localization. Propagation Review Root cause: NO FAILURE CORE Sources checked: MGI:MGI:1915008 · mouse Aadac SUPPORTS TRANSFER PANTHER:PTN002745055 · AADAC family tree node SUPPORTS TRANSFER UniProtKB:P22760 · human AADAC recipient SUPPORTS TRANSFER Self-reference: the target is its own IBD seed, which is expected rather than circular -- its own IDA/TAS annotation to this term is one of the descendant evidences behind the IBD. The IBA then asserts the additional claim that the function is inherited rather than lineage-specific. Supporting Evidence: PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. Flutamide hydrolase activity was highly detected in human liver microsomes (K(m), 794 +/- 83 microM; V(max), 1.1 +/- 0.0 nmol/min/mg protein), whereas the activity was extremely low in human liver cytosol. |
| GO:0017171 serine hydrolase activity | IBA GO_REF:0000033 | ACCEPT | Summary: AADAC is a catalytic-serine esterase/hydrolase. The family inference agrees with the conserved alpha/beta-hydrolase catalytic machinery and extensive human serine-esterase activity data. Reason: Serine hydrolase activity is a core mechanistic description of AADAC rather than an over-specific substrate claim. The human UniProt record explicitly reports serine esterase activity and the Ser-Asp-His catalytic residues, while primary human assays show inhibition by serine-esterase inhibitors. Propagation Review Root cause: NO FAILURE CORE Sources checked: MGI:MGI:1915008 · mouse Aadac SUPPORTS TRANSFER PANTHER:PTN002745055 · AADAC family tree node SUPPORTS TRANSFER UniProtKB:P22760 · human AADAC recipient SUPPORTS TRANSFER Self-reference: the target is its own IBD seed, which is expected rather than circular -- its own IDA annotation to this term is one of the descendant evidences behind the IBD. The IBA then asserts the additional claim that the function is inherited rather than lineage-specific. Supporting Evidence: file:human/AADAC/AADAC-uniprot.txt Displays serine esterase activity in liver. PMID:19339378 The same inhibition pattern was obtained with the recombinant AADAC. |
| GO:0005789 endoplasmic reticulum membrane | IEA GO_REF:0000120 | ACCEPT | Summary: Electronic orthology and UniProt subcellular-location mapping place AADAC in the endoplasmic reticulum membrane, consistent with direct human microsomal evidence. Reason: This is the correct specific membrane compartment. Human AADAC is a single-pass ER membrane enzyme and its hydrolase activity is strongly enriched in microsomes rather than cytosol (PMID:19339378). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER UniProtKB-SubCell:SL-0097 · endoplasmic reticulum SUPPORTS TRANSFER Supporting Evidence: PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. |
| GO:0008126 acetylesterase activity | IEA GO_REF:0000120 | ACCEPT | Summary: AADAC hydrolyzes diverse acetic esters, including cholesterol acetate, rifamycins, eslicarbazepine acetate, and abiraterone acetate. The electronic reaction mapping is consistent with direct human biochemical evidence. Reason: Acetylesterase activity is a core substrate-class activity of AADAC. Direct assays of recombinant human AADAC and human microsomes independently establish hydrolysis of multiple acetyl-ester drugs, while human AADAC also complements sterol-acetate turnover in yeast (PMID:18034159). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER RHEA:12957 · acetyl-ester hydrolysis reaction SUPPORTS TRANSFER EC:3.1.1.6 · acetylesterase SUPPORTS TRANSFER Supporting Evidence: PMID:18034159 The observation that expression of the human homologue of SAY1 , AADAC, in yeast rescues the sterol acetate accumulation phenotype of say1 Δ-mutant cells indicates that the human aryl acetamide deacetylase acts on cholesterol acetate PMID:33446525 Eslicarbazepine acetate was efficiently hydrolyzed by human intestinal and liver microsomes and recombinant human AADAC. |
| GO:0016020 membrane | IEA GO_REF:0000002 | MODIFY | Summary: The InterPro-derived membrane annotation is true but unnecessarily generic for a protein known to be an endoplasmic-reticulum membrane enzyme. Reason: Replace the root-level membrane term with GO:0005789 endoplasmic reticulum membrane. Direct human work localizes AADAC to the ER and human liver microsomes (PMID:19339378), so the more specific compartment is justified. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: GRANULARITY MISMATCH Sources checked: InterPro:IPR017157 · arylacetamide deacetylase family SUPPORTS TRANSFER Family membership supports membrane association but the mapped GO term is too broad. Proposed replacements: endoplasmic reticulum membrane Supporting Evidence: PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. |
| GO:0016787 hydrolase activity | IEA GO_REF:0000120 | MODIFY | Summary: Generic hydrolase activity is correct but loses the experimentally established carboxylic-ester bond class of a substantial part of AADAC catalysis. Reason: Replace GO:0016787 with GO:0052689 carboxylic ester hydrolase activity. Recombinant human AADAC hydrolyzes fluorescein diacetate and multiple acetyl-ester drugs; the replacement remains broad enough to coexist with its separate C-N hydrolase activity (PMID:26164127). This replacement consolidates with the existing GO:0052689 IEA tuple rather than proposing a second independent annotation. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: GRANULARITY MISMATCH Sources checked: ARBA:ARBA00026276 · UniProt automatic annotation rule SUPPORTS TRANSFER Supports hydrolase activity but maps only to an uninformative parent term. InterPro:IPR013094 · alpha/beta hydrolase fold-3 domain SUPPORTS TRANSFER Supports the hydrolase fold; human substrate data justify the more specific replacement. Proposed replacements: carboxylic ester hydrolase activity Supporting Evidence: PMID:26164127 Recombinant AADAC catalyzed the hydrolysis of fluorescein diacetate, N-monoacetyldapsone, and propanil, which possess notably small acyl moieties |
| GO:0047416 arylalkyl acylamidase activity | IEA GO_REF:0000120 | ACCEPT | Summary: AADAC directly deacetylates arylalkyl acylamides, including N-acetylserotonin, melatonin, and N-acetyltryptamine. The electronic reaction mapping is corroborated by recombinant-human enzyme assays. Reason: This is a specific core molecular function, not merely a name-based inference. PMID:37002641 reports conversion of N-acetylserotonin to serotonin and related reactions by recombinant human AADAC. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER RHEA:10352 · arylalkyl acylamide hydrolysis reaction SUPPORTS TRANSFER RHEA:67196 · AADAC substrate reaction SUPPORTS TRANSFER RHEA:67204 · AADAC substrate reaction SUPPORTS TRANSFER RHEA:67208 · AADAC substrate reaction SUPPORTS TRANSFER EC:3.5.1.76 · arylalkyl acylamidase SUPPORTS TRANSFER Supporting Evidence: PMID:37002641 Both human and rodent recombinant AADAC proteins can deacetylate NAS in vitro, although the human AADAC shows markedly higher activity compared with rodent enzyme. |
| GO:0052689 carboxylic ester hydrolase activity | IEA GO_REF:0000002 | ACCEPT | Summary: AADAC hydrolyzes carboxylic ester bonds in endogenous sterol acetate and several acetyl-ester drugs. The InterPro mapping is consistent with direct human evidence. Reason: Carboxylic ester hydrolase activity is a valid core parent activity for the acetylesterase and diacylglycerol-lipase reactions. Human AADAC rescues cholesterol acetate turnover in yeast and recombinant human AADAC hydrolyzes eslicarbazepine acetate (PMID:18034159; PMID:33446525). Propagation Review Root cause: NO FAILURE CORE Sources checked: InterPro:IPR017157 · arylacetamide deacetylase family SUPPORTS TRANSFER Supporting Evidence: PMID:33446525 Collectively, we found that eslicarbazepine acetate is specifically and efficiently hydrolyzed by human AADAC |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | MARK AS OVER ANNOTATED | Summary: The HuRI screen reports a binary AADAC-APPBP2 interaction, but the root-level protein binding term provides no mechanistic description of AADAC function. Reason: Retain the underlying high-throughput interaction as an observation, but do not treat generic protein binding as a core molecular function. PMID:32296183 is a proteome-scale binary-interactome map and the AADAC-APPBP2 pair has no demonstrated catalytic or physiological role here. Supporting Evidence: PMID:32296183 Here we present a human 'all-by-all' reference interactome map of human binary protein interactions, or 'HuRI'. |
| GO:0016810 hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds | IEA GO_REF:0000107 | ACCEPT | Summary: AADAC hydrolyzes non-peptide carbon-nitrogen bonds in arylacetamides such as flutamide, ketoconazole, indiplon, propanil, and N-monoacetyldapsone. The mouse orthology inference is independently corroborated by human assays. Reason: This is a correct core bond-class activity. Recombinant human AADAC directly hydrolyzes small-acyl amide substrates, so the Ensembl orthology transfer is not being accepted solely on sequence similarity (PMID:26164127; PMID:27422753). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER Supporting Evidence: PMID:26164127 Recombinant AADAC catalyzed the hydrolysis of fluorescein diacetate, N-monoacetyldapsone, and propanil, which possess notably small acyl moieties |
| GO:0017171 serine hydrolase activity | IEA GO_REF:0000107 | ACCEPT | Summary: Electronic orthology inference of serine hydrolase activity agrees with the human enzyme's catalytic-serine esterase mechanism and inhibitor profile. Reason: Serine hydrolase activity is a conserved core property of AADAC. The human record reports serine esterase activity, and recombinant-human/microsomal hydrolase activity is strongly inhibited by diisopropylphosphorofluoride and eserine (PMID:19339378). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER Supporting Evidence: file:human/AADAC/AADAC-uniprot.txt Displays serine esterase activity in liver. |
| GO:0046340 diacylglycerol catabolic process | IEA GO_REF:0000107 | ACCEPT | Summary: Mouse Aadac experimentally hydrolyzes endogenously synthesized diacylglycerol, supporting conserved involvement of human AADAC in diacylglycerol catabolism. Reason: The orthology inference is biologically coherent and has direct donor evidence. Human cellular loss/re-expression experiments also connect AADAC to neutral-lipid lipolysis, although exact purified-human DAG kinetics remain unmeasured (PMID:19654421; PMID:23542347). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER Supporting Evidence: PMID:19654421 Based on our findings, we propose a model where AADA hydrolyzes DG synthesized within the ER compartment. |
| GO:0120516 diacylglycerol lipase activity | IEA GO_REF:0000120 | ACCEPT | Summary: UniProt reaction mapping assigns diacylglycerol lipase activity from the mouse ortholog and Rhea reaction; direct mouse experiments report hydrolysis of endogenously synthesized diacylglycerol. Reason: The exact reaction is experimentally supported in the close mouse ortholog and is consistent with human AADAC-dependent cellular neutral-lipid lipolysis. Acceptance does not imply that purified-human DAG kinetic parameters are available. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER ensembl:ENSMUSP00000029325 · mouse Aadac protein SUPPORTS TRANSFER RHEA:32731 · diacylglycerol lipase reaction SUPPORTS TRANSFER Supporting Evidence: PMID:19654421 Although AADA shares similarity with HSL and like HSL showed hydrolytic activity toward endogenously synthesized DG |
| GO:0006805 xenobiotic metabolic process | IDA PMID:27422753 Human arylacetamide deacetylase hydrolyzes ketoconazole to t... | ACCEPT | Summary: Human AADAC hydrolyzes the xenobiotic ketoconazole to N-deacetyl ketoconazole, directly placing the enzyme in xenobiotic metabolism. Reason: PMID:27422753 uses human liver microsomes, recombinant enzymes, AADAC overexpression, and inhibition to assign ketoconazole hydrolysis to AADAC. A xenobiotic metabolic process annotation describes chemical transformation and does not imply that the transformation is protective. Supporting Evidence: PMID:27422753 Kinetic analysis and inhibition studies using human liver microsomes (HLM) and recombinant enzymes revealed that human arylacetamide deacetylase (AADAC) is responsible for KC hydrolysis to form DAK |
| GO:0008126 acetylesterase activity | IDA PMID:18034159 An acetylation/deacetylation cycle controls the export of st... | ACCEPT | Summary: Human AADAC hydrolyzes cholesterol acetate when expressed in yeast, rescuing the sterol-acetate accumulation phenotype of a SAY1-deficient strain. Reason: This heterologous complementation directly supports acetylesterase activity of the human protein toward a sterol acetate. It is strong evidence for the molecular activity even though it does not by itself establish a physiological human sterol pathway. Supporting Evidence: PMID:18034159 The observation that expression of the human homologue of SAY1 , AADAC, in yeast rescues the sterol acetate accumulation phenotype of say1 Δ-mutant cells indicates that the human aryl acetamide deacetylase acts on cholesterol acetate |
| GO:0008126 acetylesterase activity | IDA PMID:21856291 Human arylacetamide deacetylase is responsible for deacetyla... | ACCEPT | Summary: Recombinant human AADAC deacetylates rifampicin, rifabutin, and rifapentine to their 25-deacetylated metabolites, and human liver-microsome kinetics support the same enzyme assignment. Reason: The three rifamycin reactions are direct acetic-ester hydrolyses and fit GO:0008126. PMID:21856291 directly compares recombinant AADAC with human liver microsomes and excludes the major human carboxylesterases in this assay. Supporting Evidence: PMID:21856291 In this study, we found that recombinant human arylacetamide deacetylase (AADAC) could efficiently deacetylate rifamycins, whereas human carboxylesterases, which are enzymes responsible for the hydrolysis of many prodrugs, showed no activity. |
| GO:0008126 acetylesterase activity | IDA PMID:33446525 Role of Human Arylacetamide Deacetylase (AADAC) on Hydrolysi... | ACCEPT | Summary: Recombinant human AADAC and human intestinal and liver microsomes hydrolyze the acetyl-ester prodrug eslicarbazepine acetate to eslicarbazepine. Reason: The paper provides direct recombinant-enzyme activity, inhibitor sensitivity, correlation with human microsomal AADAC abundance, and functional effects of AADAC variants. This is strong evidence for acetylesterase activity. Supporting Evidence: PMID:33446525 Eslicarbazepine acetate was efficiently hydrolyzed by human intestinal and liver microsomes and recombinant human AADAC. |
| GO:0008126 acetylesterase activity | IDA PMID:34450168 Arylacetamide deacetylase as a determinant of the hydrolysis... | ACCEPT | Summary: Human recombinant AADAC and intestinal/liver microsomes hydrolyze abiraterone acetate to abiraterone; mouse knockout pharmacokinetics provide orthogonal in vivo support. Reason: The molecular-function term is correct, but the live QuickGO annotation search for UniProtKB:P22760 (accessed 2026-08-08) shows an extension that needs repair: it records has_input CHEBI:68642 (abiraterone), whereas the tested substrate is abiraterone acetate (CHEBI:68639) and abiraterone is the deacetylated product. This extension error does not invalidate the activity itself. Supporting Evidence: PMID:34450168 Abiraterone acetate hydrolase activity was measured using human intestinal (HIM) and liver microsomes (HLM) as well as recombinant AADAC. |
| GO:0016125 sterol metabolic process | IDA PMID:18034159 An acetylation/deacetylation cycle controls the export of st... | KEEP AS NON CORE | Summary: Human AADAC can hydrolyze cholesterol acetate in a yeast complementation system, but evidence that this reaction materially contributes to sterol metabolism in human cells or tissues is lacking. Reason: The annotation is biologically defensible because human AADAC rescues sterol-acetate accumulation in say1-deficient yeast. However, the same study reports no detectable pregnenolone acetylation in HepG2 cells and does not establish a native human sterol-acetylation cycle, so this remains a contextual/non-core process assignment. Supporting Evidence: PMID:18034159 The observation that expression of the human homologue of SAY1 , AADAC, in yeast rescues the sterol acetate accumulation phenotype of say1 Δ-mutant cells indicates that the human aryl acetamide deacetylase acts on cholesterol acetate PMID:18034159 Incubation of HepG2 hepatoma cells with radiolabeled pregnenolone, however, did not reveal any conversion to pregnenolone acetate or export of modified pregnenolone into the culture media |
| GO:0016810 hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds | IDA PMID:21856291 Human arylacetamide deacetylase is responsible for deacetyla... | ACCEPT | Summary: This experimental tuple is for flutamide amide hydrolysis, not for the rifamycin O-deacetylation foregrounded by the paper title. The live QuickGO annotation search for UniProtKB:P22760 (accessed 2026-08-08) disambiguates it with has_input CHEBI:5132 (flutamide). Reason: Flutamide hydrolysis cleaves a non-peptide C-N bond and is a directly established human AADAC activity. Although the local PMID:21856291 cache is abstract-only and foregrounds rifamycins, the curator's live flutamide extension should not be overruled from incomplete text; independent recombinant-human evidence in PMID:19339378 directly corroborates the term. Supporting Evidence: PMID:19339378 In the present study, we found that human arylacetamide deacetylase (AADAC) efficiently hydrolyzed flutamide using recombinant AADAC expressed in COS7 cells. |
| GO:0016810 hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds | IDA PMID:24464802 Indiplon is hydrolyzed by arylacetamide deacetylase in human... | ACCEPT | Summary: Recombinant human AADAC hydrolyzes the acetamide drug indiplon to deacetylindiplon, supporting non-peptide carbon-nitrogen hydrolase activity. Reason: Human liver microsomal activity, recombinant-enzyme activity, inhibitor profiling, cross-substrate correlations, and a low-activity AADAC genotype all converge on AADAC as the indiplon hydrolase (PMID:24464802). Supporting Evidence: PMID:24464802 Recombinant AADAC showed a high level of indiplon hydrolase activity, whereas recombinant carboxylesterase 1 (CES1) and 2 (CES2) showed marginal activity. |
| GO:0016810 hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds | IDA PMID:26164127 Comparison of substrate specificity among human arylacetamid... | ACCEPT | Summary: Recombinant human AADAC hydrolyzes the small-acyl amides N-monoacetyldapsone and propanil, directly demonstrating non-peptide C-N hydrolase activity. Reason: The substrate-specificity panel tested recombinant enzymes and distinguishes AADAC from CES1/CES2. The two amide substrates in the live annotation extensions support this bond-class term directly (PMID:26164127). Supporting Evidence: PMID:26164127 Recombinant AADAC catalyzed the hydrolysis of fluorescein diacetate, N-monoacetyldapsone, and propanil, which possess notably small acyl moieties |
| GO:0016810 hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds | IDA PMID:27422753 Human arylacetamide deacetylase hydrolyzes ketoconazole to t... | ACCEPT | Summary: Human AADAC hydrolyzes the N-acetyl group of ketoconazole to form N-deacetyl ketoconazole, a direct non-peptide C-N hydrolysis reaction. Reason: Recombinant-enzyme, human-liver-microsome, overexpression, and inhibitor studies assign ketoconazole hydrolysis to AADAC. The substrate-specific extension and reaction chemistry fit GO:0016810 (PMID:27422753). Supporting Evidence: PMID:27422753 Kinetic analysis and inhibition studies using human liver microsomes (HLM) and recombinant enzymes revealed that human arylacetamide deacetylase (AADAC) is responsible for KC hydrolysis to form DAK |
| GO:0098754 detoxification | IDA PMID:27422753 Human arylacetamide deacetylase hydrolyzes ketoconazole to t... | REMOVE | Summary: The cited ketoconazole reaction is metabolic activation rather than detoxification: AADAC produces N-deacetyl ketoconazole and thereby triggers hepatocellular toxicity. Reason: GO:0098754 requires a process that reduces or removes toxicity. PMID:27422753 shows the opposite direction: AADAC expression makes ketoconazole cytotoxic, while an AADAC inhibitor attenuates toxicity. The xenobiotic-metabolism annotation should be retained, but detoxification is contradicted by the cited experiment. Supporting Evidence: PMID:27422753 Overexpression of AADAC in HepaRG cells with an adenovirus expression system elicited the cytotoxicity of KC. PMID:27422753 In conclusion, the present study demonstrated that human AADAC hydrolyzes KC to trigger hepatocellular toxicity. |
| GO:0047416 arylalkyl acylamidase activity | IDA PMID:37002641 In vitro deacetylation of N-acetylserotonin by arylacetamide... | ACCEPT | Summary: Recombinant human AADAC deacetylates N-acetylserotonin to serotonin and also acts on melatonin and N-acetyltryptamine in vitro. Reason: These directly assayed arylalkyl acylamide reactions match GO:0047416. The live QuickGO annotation search for UniProtKB:P22760 (accessed 2026-08-08) nevertheless shows a polarity error: N-acetylserotonin (CHEBI:17697) is marked has_output even though it is the input substrate; that extension should be corrected without rejecting the molecular-function term. Supporting Evidence: PMID:37002641 During our search for mammalian enzymes capable of metabolizing N-acetylserotonin (NAS), AADAC was identified as having the ability to convert NAS to serotonin. |
| GO:0046340 diacylglycerol catabolic process | ISS GO_REF:0000024 | ACCEPT | Summary: Sequence-similarity transfer from mouse Aadac is supported by direct donor evidence for hydrolysis of endogenously synthesized diacylglycerol and by complementary human cellular neutral-lipid lipolysis data. Reason: Q99PG0 is a close ortholog with experimental support for the exact lipid process (PMID:19654421). Human AADAC loss/re-expression changes cellular triacylglycerol lipolysis (PMID:23542347), so this is a defensible conserved core process while purified-human DAG kinetics remain a stated gap. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER Close ortholog with direct endogenous-diacylglycerol hydrolysis evidence. Supporting Evidence: PMID:19654421 Based on our findings, we propose a model where AADA hydrolyzes DG synthesized within the ER compartment. |
| GO:0120516 diacylglycerol lipase activity | ISS GO_REF:0000024 | ACCEPT | Summary: Mouse Aadac directly hydrolyzes endogenously synthesized diacylglycerol, supporting transfer of diacylglycerol lipase activity to human AADAC. Reason: The donor is the close mouse ortholog rather than a distant paralog, and the exact activity is experimentally supported in the donor (PMID:19654421). Human cellular work corroborates a conserved lipid-lipolysis role without resolving purified-human DAG kinetics (PMID:23542347). Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:Q99PG0 · mouse Aadac SUPPORTS TRANSFER Close ortholog with direct endogenous-diacylglycerol hydrolysis evidence. Supporting Evidence: PMID:19654421 Although AADA shares similarity with HSL and like HSL showed hydrolytic activity toward endogenously synthesized DG |
| GO:0019213 deacetylase activity | IDA PMID:2043131 Purification and characterization of a human liver arylaceta... | ACCEPT | Summary: A human liver microsomal enzyme was purified on the basis of 2-acetylaminofluorene deacetylation and identified as AADAC. Reason: The purification, catalytic assay, peptide characterization, and specific antibodies directly establish deacetylase activity of the human liver protein. GO:0019213 appropriately captures the shared removal of acetyl groups across AADAC's ester and amide substrates. Supporting Evidence: PMID:2043131 An acetylaminofluorene deacetylase was purified 90 fold from human liver microsomes by PEG-fractionation, anion exchange and hydrophobic interaction chromatography. |
| GO:0019213 deacetylase activity | IDA PMID:8063807 Human liver arylacetamide deacetylase. Molecular cloning of ... | ACCEPT | Summary: Molecular cloning linked the purified human liver arylacetamide deacetylase to a 400-residue microsomal esterase involved in arylamine procarcinogen activation. Reason: The paper connects the cDNA to tryptic peptides from the purified human liver enzyme and describes the deacetylating hydrolytic reaction. This is direct support for the broad deacetylase activity term. Supporting Evidence: PMID:8063807 The cDNA was confirmed to be that for DAC in tryptic peptides from the purified human liver protein. |
| GO:0006805 xenobiotic metabolic process | TAS Reactome:R-HSA-211945 | ACCEPT | Summary: Reactome places AADAC-mediated ester and amide hydrolysis in Phase I functionalization of xenobiotic compounds. Reason: This broad process is appropriate for an enzyme that hydrolyzes flutamide, phenacetin, rifamycins, ketoconazole, and multiple prodrugs. Phase I functionalization can either facilitate elimination or generate a more reactive toxic metabolite, so acceptance is compatible with removal of the distinct detoxification annotation. |
| GO:0019213 deacetylase activity | TAS Reactome:R-HSA-5689000 | ACCEPT | Summary: The Reactome reaction assigns AADAC-catalyzed phenacetin deacetylation to p-phenetidine, a well-established human AADAC drug-hydrolysis activity. Reason: The molecular-function assertion is correct and consistent with the broader human deacetylase literature. Reactome should nevertheless revise its reaction geometry: it models phenacetin, water, and products in cytosol even though AADAC is an ER membrane protein with a lumen-facing catalytic domain. That compartment issue does not negate the enzyme activity. Supporting Evidence: Reactome:R-HSA-5689000 AADAC hydrolyses PHEN to the p-phenetidine metabolite which is a nephrotoxicant |
| GO:0005789 endoplasmic reticulum membrane | TAS Reactome:R-HSA-5689000 | ACCEPT | Summary: AADAC is an integral single-pass endoplasmic-reticulum membrane protein; Reactome's catalyst placement at the ER membrane agrees with direct human localization data. Reason: The protein-level location is correct. Reactome's associated phenacetin reaction incorrectly places its small-molecule participants in cytosol rather than the lumen-facing catalytic environment, but that model defect should not remove the independently supported ER-membrane annotation. Supporting Evidence: PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. |
| GO:0005789 endoplasmic reticulum membrane | IDA PMID:19339378 Human arylacetamide deacetylase is a principal enzyme in flu... | ACCEPT | Summary: Recombinant-human and tissue-fractionation evidence localizes AADAC to the endoplasmic reticulum and human liver microsomal fraction. Reason: PMID:19339378 directly shows recombinant AADAC activity, ER expression, strong microsomal activity, and very low cytosolic activity. This is a core cellular location for both drug and lipid hydrolysis. Supporting Evidence: PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. Flutamide hydrolase activity was highly detected in human liver microsomes (K(m), 794 +/- 83 microM; V(max), 1.1 +/- 0.0 nmol/min/mg protein), whereas the activity was extremely low in human liver cytosol. |
| GO:0017171 serine hydrolase activity | IDA PMID:17936933 Human carboxylesterases and their role in xenobiotic and end... | REMOVE | Summary: Serine hydrolase activity is correct for AADAC, but PMID:17936933 is a review of human carboxylesterases rather than a primary-research report and therefore cannot support an IDA evidence code. Reason: Remove this source-specific IDA tuple on the narrow evidence-code ground that the cited article is explicitly classified as a review and cannot itself report a direct assay. This does not reject either the review's relevance to AADAC or the function: independent primary human work and other retained annotations establish serine-hydrolase activity (PMID:19339378). Supporting Evidence: file:human/AADAC/AADAC-uniprot.txt Displays serine esterase activity in liver. PMID:19339378 The same inhibition pattern was obtained with the recombinant AADAC. |
| GO:0016298 lipase activity | TAS PMID:11481320 Characterization of the rodent genes for arylacetamide deace... | MODIFY | Summary: The 2001 rodent-gene paper proposed AADAC as a microsomal lipase from homology and expression patterns, but later work resolves its endogenous neutral-lipid activity more specifically as diacylglycerol lipase activity. Reason: Replace generic GO:0016298 with GO:0120516 diacylglycerol lipase activity. The cited PMID:11481320 provides only a putative rodent lipase hypothesis; subsequent mouse-ortholog experiments report hydrolysis of endogenously synthesized DG (PMID:19654421), and human AADAC perturbation changes cellular TG lipolysis (PMID:23542347). Proposed replacements: diacylglycerol lipase activity Supporting Evidence: PMID:11481320 The homology of AADA with hormone-sensitive lipase and the tissue distribution of AADA are consistent with the view that AADA plays a role in promoting the mobilization of lipids from intracellular stores and in the liver for assembling VLDL. PMID:19654421 Although AADA shares similarity with HSL and like HSL showed hydrolytic activity toward endogenously synthesized DG |
| GO:0005789 endoplasmic reticulum membrane | IDA PMID:10318829 Targeting proteins to the lumen of endoplasmic reticulum usi... | ACCEPT | Summary: The cached abstract characterizes the legacy 50-kDa esterase/E3 as an intrinsic type-II ER membrane protein with a lumen-facing C terminus; direct human AADAC work independently corroborates the ER location. Reason: The abstract does not identify the P22760 accession or species of every construct, so the legacy E3 identity is not overinterpreted. The experimental location itself is concordant with direct human AADAC microsomal/ER evidence in PMID:19339378, making the annotation biologically secure despite imperfect source provenance. Supporting Evidence: PMID:10318829 Both are type II membrane proteins with the C terminus projecting into the lumen of the ER. PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. |
| GO:0005789 endoplasmic reticulum membrane | IDA PMID:15152005 Appropriate function of 11beta-hydroxysteroid dehydrogenase ... | UNDECIDED | Summary: The abstract maps N-terminal topological determinants in E3/50-kDa esterase, but the local cache lacks full text and does not establish the species of the tested E3 constructs well enough to adjudicate a human AADAC IDA tuple. Reason: Use UNDECIDED because the relevant full text is unavailable in the repository and organism attribution cannot be safely inferred from the abstract. The abstract's Lys-4 and Asp-25 E3 determinants are compatible with human P22760 numbering, while multiple retained annotations and PMID:19339378 independently establish the human ER-membrane location. Supporting Evidence: PMID:15152005 Previous studies revealed that the luminal orientation of 11beta-HSD1 and 50-kDa esterase/arylacetamide deacetylase (E3) is determined by their highly similar N-terminal transmembrane domains. PMID:19339378 AADAC is specifically expressed in the endoplasmic reticulum. |
| GO:0019213 deacetylase activity | IDA PMID:9665742 Determination of lumenal orientation of microsomal 50-kDa es... | UNDECIDED | Summary: The abstract studies topology and post-translational structure of a legacy 50-kDa esterase/N-deacetylase, but does not expose the organism or the catalytic assay underlying the human deacetylase IDA tuple. Reason: Use UNDECIDED because the repository cache is abstract-only. The reported Asn-77/Asn-281 sites could reflect mature-protein numbering relative to human Asn-78/Asn-282, while its Cys-115/Cys-340 bond aligns with the human UniProt 116..340 disulfide at only one end; these offsets are not a clean ortholog signature, and the abstract does not permit a reliable species assignment. Direct human PMIDs 2043131 and 8063807 independently retain the correct deacetylase function. Supporting Evidence: PMID:2043131 This report thus describes the first arylacetamide deacetylase in human liver. PMID:8063807 Microsomal arylacetamide deacetylase (DAC) competes against the activity of cytosolic arylamine N-acetyltransferase |
| GO:0003824 catalytic activity | TAS PMID:8063807 Human liver arylacetamide deacetylase. Molecular cloning of ... | MODIFY | Summary: Root-level catalytic activity is true but uninformative for a cloned human enzyme whose deacetylase activity and substrate class were already established. Reason: Replace GO:0003824 with GO:0019213 deacetylase activity. PMID:8063807 identifies the cloned human liver protein as the microsomal arylacetamide deacetylase involved in the hydrolytic activation of arylamine carcinogens, supporting the specific parent activity rather than the ontology root. Proposed replacements: deacetylase activity Supporting Evidence: PMID:8063807 This cDNA provides an important tool to study deacetylation and its effects on the metabolic activation of arylamine and heterocyclic amine carcinogens. |
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Download this section (compressed HTML)Q: Does full-length human AADAC directly hydrolyze diacylglycerol, and what are its preferred DAG molecular species and kinetic parameters relative to triacylglycerol, cholesteryl ester, and small-acyl xenobiotic substrates?
Q: How do xenobiotics and membrane lipids reach the lumen-facing AADAC catalytic region: by passive partitioning, intramembrane access, lipid translocation, or a dedicated ER transport mechanism?
Q: In native human liver and intestine, how much does AADAC contribute to DAG and triacylglycerol flux, lipid-droplet mobilization, and VLDL production across nutritional and disease states relative to other ER hydrolases?
Q: Are cholesterol acetate, N-acetylserotonin, melatonin, or N-acetyltryptamine physiologically important AADAC substrates in human tissues at endogenous concentrations, or only context-specific in-vitro substrates?
Experiment: Purify full-length human AADAC and reconstitute it in oriented ER-like proteoliposomes. Compare wild-type protein with a catalytic-serine mutant across a panel of defined DAG species, triacylglycerols, cholesteryl esters, and established small-acyl drug substrates; quantify monoacylglycerol and fatty-acid products by targeted mass spectrometry and determine kinetic parameters.
Hypothesis: Human AADAC directly hydrolyzes a restricted set of diacylglycerol molecular species through its serine-hydrolase active site.
Type: oriented-membrane recombinant enzymology and lipidomics
Experiment: Perform AADAC knockout or CRISPRi followed by wild-type versus catalytic-dead rescue in primary human hepatocytes and intestinal organoids. Use stable-isotope tracing and quantitative lipidomics to measure DAG, triacylglycerol, monoacylglycerol, lipid-droplet turnover, and secreted lipoproteins, with CES2 perturbation as a pathway-specific comparator.
Hypothesis: AADAC catalytic activity makes a measurable, tissue-dependent contribution to human hepatic and intestinal neutral-lipid flux and VLDL assembly.
Type: human-cell genetic rescue and stable-isotope lipid flux analysis
Experiment: Use sealed microsomes or oriented proteoliposomes with independently verified AADAC topology, selectively deliver membrane-permeant and membrane-impermeant substrate analogs to either face, and combine protease protection with transport or lipid-flip inhibition to distinguish passive partitioning from carrier-dependent access.
Hypothesis: Substrate access to AADAC depends on presentation to the ER-lumenal face rather than unrestricted exposure from the cytosol.
Type: sided ER-membrane substrate-access assay
Experiment: In human hepatic, intestinal, and pineal-relevant cell models with verified endogenous AADAC, use knockout and wild-type versus catalytic-dead rescue together with isotope-labeled cholesterol acetate, N-acetylserotonin, melatonin, and N-acetyltryptamine at physiological concentrations. Quantify substrate-product flux and compare it with established xenobiotic deacetylation controls.
Hypothesis: The reported sterol-acetate and pineal-indole reactions contribute measurable flux in only selected human tissues or physiological states.
Type: tissue-context genetic rescue and targeted metabolite tracing
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