CAD

Summary

CAD (carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase, HGNC:1424) is a protein-coding gene on chromosome 2p23.3, encoding Multifunctional protein CAD (P27708). Multifunctional protein that encodes the first 3 enzymatic activities of the de novo pyrimidine pathway: carbamoylphosphate synthetase (CPSase; EC 6.3.5.5), aspartate transcarbamylase (ATCase; EC 2.1.3.2) and dihydroorotase (DHOase; EC 3.5.2.3).

The de novo synthesis of pyrimidine nucleotides is required for mammalian cells to proliferate. This gene encodes a trifunctional protein which is associated with the enzymatic activities of the first 3 enzymes in the 6-step pathway of pyrimidine biosynthesis: carbamoylphosphate synthetase (CPS II), aspartate transcarbamoylase, and dihydroorotase. This protein is regulated by the mitogen-activated protein kinase (MAPK) cascade, which indicates a direct link between activation of the MAPK cascade and de novo biosynthesis of pyrimidine nucleotides. Alternative splicing results in multiple transcript variants encoding different isoforms.

Source: NCBI Gene 790 — RefSeq curated summary.

At a glance

• Gene–disease (curated): developmental and epileptic encephalopathy, 50 (Definitive, ClinGen)
• GWAS associations: 1
• Clinical variants (ClinVar): 2,342 total — 62 pathogenic, 32 likely-pathogenic
• Phenotypes (HPO): 25
• Druggable target: yes
• MANE Select transcript: NM_004341

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:1424
Approved symbol	CAD
Name	carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase
Location	2p23.3
Locus type	gene with protein product
Status	Approved
Aliases	GATD4
Ensembl gene	ENSG00000084774
Ensembl biotype	protein_coding
OMIM	114010
Entrez	790

Gene structure

Transcript identifiers

Ensembl transcripts: 13 — 7 protein_coding, 4 retained_intron, 2 protein_coding_CDS_not_defined

ENST00000264705, ENST00000403525, ENST00000428460, ENST00000456311, ENST00000458503, ENST00000464159, ENST00000475695, ENST00000479002, ENST00000487239, ENST00000491461, ENST00000491891, ENST00000854433, ENST00000912693

RefSeq mRNA: 2 — MANE Select: NM_004341 NM_001306079, NM_004341

CCDS: CCDS1742, CCDS77396

Canonical transcript exons

ENST00000264705 — 44 exons

Exon	Start	End
ENSE00000732839	27217877	27218016
ENSE00000732842	27221218	27221347
ENSE00000732854	27222866	27223037
ENSE00000732858	27223563	27223748
ENSE00000732861	27223917	27224029
ENSE00000732870	27224745	27224876
ENSE00000732874	27225010	27225243
ENSE00000732878	27225705	27225926
ENSE00000732882	27226131	27226319
ENSE00000732885	27226525	27226649
ENSE00000732901	27233042	27233140
ENSE00000732910	27233626	27233808
ENSE00000732912	27234008	27234226
ENSE00000732914	27234518	27234685
ENSE00000733044	27238431	27238632
ENSE00000733057	27240265	27240361
ENSE00000733067	27241322	27241396
ENSE00000733070	27241911	27242123
ENSE00000733074	27242302	27242427
ENSE00000733078	27242620	27242775
ENSE00000733082	27242872	27242973
ENSE00000733087	27243198	27243292
ENSE00000809111	27217369	27217633
ENSE00000809117	27235536	27235640
ENSE00000809118	27236284	27236523
ENSE00000809121	27237379	27237545
ENSE00001035908	27222194	27222336
ENSE00001078281	27237718	27237882
ENSE00001078282	27236749	27236830
ENSE00001078283	27238056	27238187
ENSE00001078284	27222519	27222660
ENSE00001078287	27233312	27233536
ENSE00001078288	27224345	27224490
ENSE00001078290	27241058	27241227
ENSE00001078291	27232448	27232694
ENSE00001078293	27240911	27240955
ENSE00001078294	27239042	27239232
ENSE00001245850	27243416	27243943
ENSE00003499463	27226832	27226962
ENSE00003539197	27239331	27239471
ENSE00003567777	27231468	27231580
ENSE00003590301	27231980	27232224
ENSE00003646700	27239697	27239798
ENSE00003656422	27235245	27235427

Expression profiles

Bgee: expression breadth ubiquitous, 223 present calls, max score 90.89.

FANTOM5 (CAGE): breadth ubiquitous, TPM avg 43.3835 / max 477.6382, expressed in 1785 samples.

FANTOM5 promoters (7 alternative TSS)

Promoter ID	TPM avg	Samples expressed
19333	24.8228	1718
19334	7.5937	1525
19335	7.4323	1694
19336	2.1368	1136
19332	1.0878	652
19339	0.2047	105
19337	0.1055	42

Top tissues by expression

282 total, by Bgee expression score (0-100, higher = more expressed):

Tissue	Anatomy ID	Expression score	Quality
body of uterus	UBERON:0009853	90.89	gold quality
stromal cell of endometrium	CL:0002255	90.68	gold quality
right lobe of liver	UBERON:0001114	90.52	gold quality
primordial germ cell in gonad	CL:0000670 ∩ UBERON:0000991	90.46	gold quality
ventricular zone	UBERON:0003053	90.32	gold quality
right testis	UBERON:0004534	90.18	gold quality
right uterine tube	UBERON:0001302	90.15	gold quality
left testis	UBERON:0004533	90.08	gold quality
right ovary	UBERON:0002118	90.06	gold quality
mucosa of transverse colon	UBERON:0004991	90.03	gold quality
lower esophagus mucosa	UBERON:0035834	89.98	gold quality
right lobe of thyroid gland	UBERON:0001119	89.84	gold quality
left ovary	UBERON:0002119	89.78	gold quality
ectocervix	UBERON:0012249	89.51	gold quality
left uterine tube	UBERON:0001303	89.26	gold quality
adenohypophysis	UBERON:0002196	89.23	gold quality
apex of heart	UBERON:0002098	88.93	gold quality
tibial nerve	UBERON:0001323	88.85	gold quality
right hemisphere of cerebellum	UBERON:0014890	88.85	gold quality
metanephros cortex	UBERON:0010533	88.82	gold quality
pituitary gland	UBERON:0000007	88.70	gold quality
endocervix	UBERON:0000458	88.64	gold quality
endothelial cell	CL:0000115	88.57	silver quality
left lobe of thyroid gland	UBERON:0001120	88.19	gold quality
cerebellar hemisphere	UBERON:0002245	87.90	gold quality
cerebellar cortex	UBERON:0002129	87.73	gold quality
muscle layer of sigmoid colon	UBERON:0035805	87.70	gold quality
body of pancreas	UBERON:0001150	87.65	gold quality
testis	UBERON:0000473	87.34	gold quality
skin of leg	UBERON:0001511	87.23	gold quality

Single-cell (SCXA)

Detected in 1 experiment(s), a significant marker in 1.

Experiment	Marker?	Max mean expression
E-ANND-3	yes	5.35

Regulation

Is transcription factor: no

miRNA regulators (miRDB)

17 targeting CAD, top 30 by miRDB confidence (max_score; target_count = how many genes the miRNA targets in total — lower means more specific):

miRNA	Max score	Avg score	miRNA target_count
HSA-MIR-486-3P	99.51	66.82	1901
HSA-MIR-4311	99.31	70.47	3041
HSA-MIR-18A-5P	99.29	71.05	806
HSA-MIR-18B-5P	99.29	71.05	806
HSA-MIR-4293	99.22	65.46	1263
HSA-MIR-4735-3P	99.14	69.85	777
HSA-MIR-4695-5P	99.06	64.87	1151
HSA-MIR-936	98.87	70.51	1124
HSA-MIR-4477A	98.83	69.75	2952
HSA-MIR-222-5P	98.75	69.17	1242
HSA-MIR-595	98.25	67.44	699
HSA-MIR-6882-3P	98.23	67.01	1119
HSA-MIR-943	97.81	64.42	694
HSA-MIR-4313	97.18	63.15	420
HSA-MIR-1913	97.07	66.20	1417
HSA-MIR-2861	95.24	65.47	1056
HSA-MIR-11400	94.03	67.12	81

Literature-anchored findings (GeneRIF, showing 23)

• the cad gene is regulated by a nonclassical ERalpha/Sp1-mediated pathway. (PMID:12746293)
• Data show that PRMT5 can be found in association with hSWI/SNF complexes and is involved in regulating the expression of carbamoyl-phosphate synthase-aspartate carbamoyltransferase-dihydroorotase. (PMID:14559996)
• the nuclear import of CAD appears to promote optimal cell growth (PMID:15890648)
• cad gene expression is repressed by hypoxia-inducible factor-1alpha, which represents a functional link between hypoxia and cell-cycle arrest. (PMID:16155188)
• hCPS_DeltaA was not able to fully assume the catalytically competent conformation, with specific activity of CP formation decreased 700-fold. (PMID:18679823)
• findings show that in prostate tumor cells, CAD fosters androgen receptor translocation into the nucleus and stimulates its transcriptional activity; in radical prostatectomy specimens, CAD expression was not correlated with proliferation markers, but a higher CAD mRNA level was associated with local tumor extension and cancer relapse (PMID:21982950)
• preliminary X-ray diffraction analysis of the dihydroorotase domain of human CAD (PMID:23143245)
• The results obtained indicate that mLST8 bridges between CAD and mTOR, and plays a role in the signaling mechanism where CAD is regulated in the mTOR pathway through the association with mLST8 (PMID:23594158)
• Recombinant aspartate carbamoyltransferase domain from the CAD enzyme complex forms homotrimers in solution. (PMID:24316846)
• These results establish CAD as a downstream effector of Rheb and suggest a possible role of Rheb in regulating de novo pyrimidine nucleotide synthesis (PMID:25422319)
• Changes in glycosylation in caused by mutations in CAD. (PMID:25678555)
• Previous study found three metal ions in huDHOase active site; in the present study, the putative third metal binding site in type II enzymes, such as StDHOase and ScDHOase, was created and identified. (PMID:26446564)
• This study showed that CAD deficiency co-occurrence of anaemia, anisopoikilocytosis, global developmental delay, and seizures. (PMID:28007989)
• Detection of the CAD-ALK gene fusion in urine tr-DNA anticipated radiological confirmation of disease progression. Analysis of plasma ctDNA identified ALK kinase mutations that emerged during treatment with the ALK inhibitor entrectinib (PMID:28368455)
• CAD dihydroorotase and aspartate transcarbamoylase domains self-assembles into dimers and trimers. (PMID:28552578)
• Charge neutralization in the active site of the catalytic trimer of aspartate transcarbamoylase promotes diverse structural changes. (PMID:28833948)
• structural evidences indicated that post-translational carbamylated Lys was not required for Znalpha binding in PaDHPase and in huDHOase. (PMID:30268498)
• The catalytic flexibile loop in the dihydrooratase domain possesses an absolutely conserved Phe-1563 residue required for catalytic activity. (PMID:30315107)
• New regulatory mechanism-based inhibitors of aspartate transcarbamoylase for potential anticancer drug development. (PMID:31967710)
• The Cellular Protein CAD is Recruited into Ebola Virus Inclusion Bodies by the Nucleoprotein NP to Facilitate Genome Replication and Transcription. (PMID:32370067)
• Uridine treatment normalizes the congenital dyserythropoietic anemia type II-like hematological phenotype in a patient with homozygous mutation in the CAD gene. (PMID:32720728)
• Deciphering CAD: Structure and function of a mega-enzymatic pyrimidine factory in health and disease. (PMID:34288185)
• Beyond genetics: Deciphering the impact of missense variants in CAD deficiency. (PMID:37540500)

Cross-species orthologs

5 orthologs

Organism	Symbol	Gene ID
danio_rerio	cad	ENSDARG00000041895
mus_musculus	Cad	ENSMUSG00000013629
rattus_norvegicus	Cad	ENSRNOG00000026474
drosophila_melanogaster	r	FBGN0003189
caenorhabditis_elegans		WBGENE00004259

Paralogs (2): CPS1 (ENSG00000021826), OTC (ENSG00000036473)

Protein

Protein identifiers

Multifunctional protein CAD — P27708 (reviewed: P27708)

Alternative names: Carbamoyl phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase

All UniProt accessions (5): P27708, F8VPD4, H7BZB3, H7C2E4, H7C3Z5

UniProt curated annotations — full annotation on UniProt →

Function. Multifunctional protein that encodes the first 3 enzymatic activities of the de novo pyrimidine pathway: carbamoylphosphate synthetase (CPSase; EC 6.3.5.5), aspartate transcarbamylase (ATCase; EC 2.1.3.2) and dihydroorotase (DHOase; EC 3.5.2.3). The CPSase-function is accomplished in 2 steps, by a glutamine-dependent amidotransferase activity (GATase) that binds and cleaves glutamine to produce ammonia, followed by an ammonium-dependent carbamoyl phosphate synthetase, which reacts with the ammonia, hydrogencarbonate and ATP to form carbamoyl phosphate. The endogenously produced carbamoyl phosphate is sequestered and channeled to the ATCase active site. ATCase then catalyzes the formation of carbamoyl-L-aspartate from L-aspartate and carbamoyl phosphate. In the last step, DHOase catalyzes the cyclization of carbamoyl aspartate to dihydroorotate.

Subunit / interactions. Homohexamer. Interacts with CIPC.

Subcellular location. Cytoplasm. Nucleus.

Post-translational modifications. Activated by MAP kinase (Erk1/2) phosphorylation just prior to the S phase of the cell cycle, when the demand for pyrimidine nucleotides is greatest, and down-regulated as the cells emerge from S phase by protein kinase A (PKA) phosphorylation. Phosphorylation at Ser-1859 by RPS6KB1 downstream of MTOR promotes oligomerization and stimulates dihydroorotase activity. Phosphorylation at Ser-1406 reduces sensitivity to feedback inhibition by UTP.

Disease relevance. Developmental and epileptic encephalopathy 50 (DEE50) [MIM:616457] A form of epileptic encephalopathy, a heterogeneous group of severe early-onset epilepsies characterized by refractory seizures, neurodevelopmental impairment, and poor prognosis. Development is normal prior to seizure onset, after which cognitive and motor delays become apparent. DEE50 is an autosomal recessive, progressive disease with onset in infancy and favorable response to treatment with oral uridine. The disease is caused by variants affecting the gene represented in this entry.

Activity regulation. Allosterically regulated and controlled by phosphorylation. 5-phosphoribose 1-diphosphate (PRPP) is an activator while UMP and UTP are inhibitors of the CPSase reaction.

Cofactor. Binds 3 Zn(2+) ions per subunit (for dihydroorotase activity). Binds 4 magnesium or manganese ions per subunit.

Induction. Transcriptionally repressed following hypoxia by HIF1A.

Pathway. Pyrimidine metabolism; UMP biosynthesis via de novo pathway; (S)-dihydroorotate from bicarbonate: step 1/3. Pyrimidine metabolism; UMP biosynthesis via de novo pathway; (S)-dihydroorotate from bicarbonate: step 2/3. Pyrimidine metabolism; UMP biosynthesis via de novo pathway; (S)-dihydroorotate from bicarbonate: step 3/3.

Miscellaneous. GATase (glutamine amidotransferase) and CPSase (carbamoyl phosphate synthase) form together the glutamine-dependent CPSase (GD-CPSase) (EC 6.3.5.5).

Similarity. In the N-terminal section; belongs to the CarA family. In the 2nd section; belongs to the CarB family. In the 3rd section; belongs to the metallo-dependent hydrolases superfamily. DHOase family. CAD subfamily. In the C-terminal section; belongs to the aspartate/ornithine carbamoyltransferase superfamily. ATCase family.

RefSeq proteins (2): NP_001293008, NP_004332* (*=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR002082	Asp_carbamoyltransf	Family
IPR002195	Dihydroorotase_CS	Conserved_site
IPR002474	CarbamoylP_synth_ssu_N	Domain
IPR005479	CPAse_ATP-bd	Domain
IPR005480	CPSase_lsu_oligo	Domain
IPR005483	CPSase_dom	Domain
IPR006130	Asp/Orn_carbamoylTrfase	Family
IPR006131	Asp_carbamoyltransf_Asp/Orn-bd	Domain
IPR006132	Asp/Orn_carbamoyltranf_P-bd	Domain
IPR006274	CarbamoylP_synth_ssu	Family
IPR006275	CPSase_lsu	Family
IPR006680	Amidohydro-rel	Domain
IPR011059	Metal-dep_hydrolase_composite	Homologous_superfamily
IPR011607	MGS-like_dom	Domain
IPR011761	ATP-grasp	Domain
IPR013815	ATP_grasp_subdomain_1	Homologous_superfamily
IPR016185	PreATP-grasp_dom_sf	Homologous_superfamily
IPR017926	GATASE	Domain
IPR029062	Class_I_gatase-like	Homologous_superfamily
IPR032466	Metal_Hydrolase	Homologous_superfamily
IPR035686	CPSase_GATase1	Domain
IPR036480	CarbP_synth_ssu_N_sf	Homologous_superfamily
IPR036897	CarbamoylP_synth_lsu_oligo_sf	Homologous_superfamily
IPR036901	Asp/Orn_carbamoylTrfase_sf	Homologous_superfamily
IPR036914	MGS-like_dom_sf	Homologous_superfamily
IPR058047	CPSase_preATP-grasp	Domain

Pfam: PF00117, PF00185, PF00988, PF01979, PF02142, PF02729, PF02786, PF02787, PF25596

Enzyme classification (BRENDA):

• EC 2.1.3.2 — aspartate carbamoyltransferase (BRENDA: 52 organisms, 33 substrates, 217 inhibitors, 81 Km, 10 kcat entries)
• EC 3.5.2.3 — dihydroorotase (BRENDA: 34 organisms, 29 substrates, 74 inhibitors, 68 Km, 27 kcat entries)
• EC 6.3.5.5 — carbamoyl-phosphate synthase (glutamine-hydrolysing) (BRENDA: 45 organisms, 73 substrates, 72 inhibitors, 194 Km, 142 kcat entries)

Substrate kinetics (BRENDA)

23 substrates with measured Km, best-characterized 15. Km ranges are aggregated across organisms/conditions.

Substrate	Km (mM)	Measurements
ATP	0.004–8.36	84
CARBAMOYL PHOSPHATE	0.004–88	33
L-ASPARTATE	0.009–44.7	33
L-GLN	0.0119–22	29
L-GLUTAMINE	0.04–18.87	25
HCO3-	0.64–32.5	16
L-DIHYDROOROTATE	0.004–0.45	15
NH4+	9.8–160	14
N-CARBAMOYL-L-ASPARTATE	0.0281–24	12
ADP	0.022–0.53	11
(S)-DIHYDROOROTATE	0.02–0.6024	9
N-(AMINOCARBONYL)-L-ASPARTIC ACID	0.285–6	7
CARBAMOYL ASPARTATE	0.51–15	6
L-CARBAMOYLASPARTATE	0.015–2.2	6
DIHYDROOROTATE	0.063–3.03	5

Catalyzed reactions (Rhea), 5 shown:

• L-glutamine + H2O = L-glutamate + NH4(+) (RHEA:15889)
• hydrogencarbonate + NH4(+) + 2 ATP = carbamoyl phosphate + 2 ADP + phosphate + 2 H(+) (RHEA:18029)
• hydrogencarbonate + L-glutamine + 2 ATP + H2O = carbamoyl phosphate + L-glutamate + 2 ADP + phosphate + 2 H(+) (RHEA:18633)
• carbamoyl phosphate + L-aspartate = N-carbamoyl-L-aspartate + phosphate + H(+) (RHEA:20013)
• (S)-dihydroorotate + H2O = N-carbamoyl-L-aspartate + H(+) (RHEA:24296)

UniProt features (198 total): binding site 70, helix 34, strand 26, mutagenesis site 15, modified residue 12, region of interest 9, turn 9, sequence conflict 6, sequence variant 5, active site 4, domain 4, compositionally biased region 2, initiator methionine 1, chain 1

Structure

Experimental structures (PDB)

55 structures, top 30 by resolution.

PDB	Method	Resolution (Å)
9FS2	X-RAY DIFFRACTION	1.12
6HFQ	X-RAY DIFFRACTION	1.15
9FS3	X-RAY DIFFRACTION	1.18
6HFP	X-RAY DIFFRACTION	1.2
6HFJ	X-RAY DIFFRACTION	1.2
9FS1	X-RAY DIFFRACTION	1.21
4C6F	X-RAY DIFFRACTION	1.26
4C6E	X-RAY DIFFRACTION	1.26
8PBM	X-RAY DIFFRACTION	1.28
4C6D	X-RAY DIFFRACTION	1.3
4C6J	X-RAY DIFFRACTION	1.3
6HFR	X-RAY DIFFRACTION	1.3
6HFS	X-RAY DIFFRACTION	1.35
4C6I	X-RAY DIFFRACTION	1.35
6HFL	X-RAY DIFFRACTION	1.35
6HFU	X-RAY DIFFRACTION	1.4
8PBT	X-RAY DIFFRACTION	1.43
6HFH	X-RAY DIFFRACTION	1.45
6HFN	X-RAY DIFFRACTION	1.45
4C6C	X-RAY DIFFRACTION	1.45
6HFK	X-RAY DIFFRACTION	1.46
6HFI	X-RAY DIFFRACTION	1.46
8PBG	X-RAY DIFFRACTION	1.46
4C6K	X-RAY DIFFRACTION	1.48
6HFE	X-RAY DIFFRACTION	1.48
8PBI	X-RAY DIFFRACTION	1.5
6HFF	X-RAY DIFFRACTION	1.51
4C6P	X-RAY DIFFRACTION	1.52
8PBP	X-RAY DIFFRACTION	1.54
8PBQ	X-RAY DIFFRACTION	1.54

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-P27708-F1	87.25	0.62

Functional residue map

Curated UniProt residues grouped by drug-discovery relevance — catalytic, ligand-binding, modification, and mutation-validated positions. Source: UniProtKB sequence features.

Catalytic / active sites (4): 252 (nucleophile; for gatase activity); 336 (for gatase activity); 338 (for gatase activity); 1686 (for dhoase activity)

Ligand- & substrate-binding residues (70): 44; 222; 224; 253; 256; 294; 296; 297; 515; 555; 561; 562 …

Post-translational modifications (12): 1884, 1900, 1938, 2, 456, 747, 1038, 1406, 1411, 1556, 1859, 1873

Mutagenesis-validated functional residues (15):

Position	Phenotype
1471	no zinc-binding and no catalytic activity.
1471	abolishes dihydroorotase activity.
1473	no zinc-binding and no catalytic activity.
1512	no change in catalytic activity.
1562	abolishes dihydroorotase activity.
1563	abolishes dihydroorotase activity.
1590	abolishes dihydroorotase activity.
1590	no catalytic activity.
1613	reduces dihydroorotase activity.
1614	abolishes dihydroorotase activity.
1637	abolishes dihydroorotase activity.
1642	11.5% of wild-type catalytic activity.
1686	abolishes dihydroorotase activity.
1690	3% of wild-type catalytic activity.
1873	abolishes pma-induced thr-456 phosphorylation.

Function

Pathways and Gene Ontology

Reactome pathways

1 pathways

ID	Pathway
R-HSA-500753	Pyrimidine biosynthesis

MSigDB gene sets: 404 (showing top): GSE45365_NK_CELL_VS_CD8A_DC_UP, GSE45365_NK_CELL_VS_BCELL_UP, GOBP_RESPONSE_TO_NITROGEN_COMPOUND, GOBP_HEPATICOBILIARY_SYSTEM_DEVELOPMENT, GOBP_NUCLEOSIDE_DIPHOSPHATE_METABOLIC_PROCESS, GOBP_RESPONSE_TO_AMINE, GOBP_RESPONSE_TO_CORTICOSTEROID, GOBP_ALPHA_AMINO_ACID_METABOLIC_PROCESS, MATTIOLI_MGUS_VS_PCL, CAIRO_PML_TARGETS_BOUND_BY_MYC_UP, MORF_CDK2, GOBP_PYRIMIDINE_NUCLEOSIDE_TRIPHOSPHATE_BIOSYNTHETIC_PROCESS, GOBP_REGENERATION, MITSIADES_RESPONSE_TO_APLIDIN_DN, GOBP_ORGANOPHOSPHATE_METABOLIC_PROCESS

GO Biological Process (25): liver development (GO:0001889), ‘de novo’ pyrimidine nucleobase biosynthetic process (GO:0006207), UDP biosynthetic process (GO:0006225), UTP biosynthetic process (GO:0006228), L-glutamine metabolic process (GO:0006541), xenobiotic metabolic process (GO:0006805), heart development (GO:0007507), female pregnancy (GO:0007565), lactation (GO:0007595), response to amine (GO:0014075), L-citrulline biosynthetic process (GO:0019240), response to caffeine (GO:0031000), animal organ regeneration (GO:0031100), response to insulin (GO:0032868), response to testosterone (GO:0033574), response to starvation (GO:0042594), ‘de novo’ UMP biosynthetic process (GO:0044205), response to cortisol (GO:0051414), cellular response to epidermal growth factor stimulus (GO:0071364), pyrimidine nucleotide biosynthetic process (GO:0006221), amino acid metabolic process (GO:0006520), phosphate-containing compound metabolic process (GO:0006796), nucleobase-containing small molecule metabolic process (GO:0055086), cellular response to xenobiotic stimulus (GO:0071466), pyrimidine-containing compound biosynthetic process (GO:0072528)

GO Molecular Function (22): aspartate carbamoyltransferase activity (GO:0004070), carbamoyl-phosphate synthase (ammonia) activity (GO:0004087), carbamoyl-phosphate synthase (glutamine-hydrolyzing) activity (GO:0004088), dihydroorotase activity (GO:0004151), glutaminase activity (GO:0004359), protein kinase activity (GO:0004672), ATP binding (GO:0005524), zinc ion binding (GO:0008270), enzyme binding (GO:0019899), identical protein binding (GO:0042802), aspartate binding (GO:0070335), nucleotide binding (GO:0000166), catalytic activity (GO:0003824), protein binding (GO:0005515), amino acid binding (GO:0016597), transferase activity (GO:0016740), carboxyl- or carbamoyltransferase activity (GO:0016743), hydrolase activity (GO:0016787), hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds (GO:0016810), hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in cyclic amides (GO:0016812), ligase activity (GO:0016874), metal ion binding (GO:0046872)

GO Cellular Component (10): nucleus (GO:0005634), cytoplasm (GO:0005737), cytosol (GO:0005829), membrane (GO:0016020), nuclear matrix (GO:0016363), protein-containing complex (GO:0032991), cell projection (GO:0042995), neuronal cell body (GO:0043025), terminal bouton (GO:0043195), extracellular exosome (GO:0070062)

Reactome top-level categories

Rollup of top-1 pathways:

Category	Pathways
Nucleotide biosynthesis	1

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
cellular anatomical structure	5
pyrimidine ribonucleotide biosynthetic process	2
animal organ development	2
response to ketone	2
protein binding	2
binding	2
catalytic activity	2
gland development	1
hepaticobiliary system development	1
pyrimidine nucleobase biosynthetic process	1
pyrimidine ribonucleoside diphosphate biosynthetic process	1
UDP metabolic process	1
pyrimidine ribonucleoside triphosphate biosynthetic process	1
UTP metabolic process	1
L-amino acid metabolic process	1
proteinogenic amino acid metabolic process	1
metabolic process	1
cellular response to xenobiotic stimulus	1
circulatory system development	1
multi-organism reproductive process	1
multi-multicellular organism process	1
body fluid secretion	1
mammary gland development	1
milk ejection reflex	1
response to nitrogen compound	1
amino acid biosynthetic process	1
non-proteinogenic amino acid biosynthetic process	1
response to purine-containing compound	1
response to alkaloid	1
regeneration	1
response to peptide hormone	1
response to lipid	1
response to stress	1
response to nutrient levels	1
UMP biosynthetic process	1
response to glucocorticoid	1
response to alcohol	1
response to epidermal growth factor	1
cellular response to growth factor stimulus	1
pyrimidine nucleotide metabolic process	1

Protein interactions and networks

STRING

2836 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
CAD	UMPS	P11172	805
CAD	PPAT	Q06203	805
CAD	DHODH	Q02127	779
CAD	CTPS1	P17812	588
CAD	GMPS	P49915	585
CAD	ASS1	P00966	584
CAD	GART	P22102	581
CAD	ADSL	P30566	568
CAD	MLH1	P40692	558
CAD	DTYMK	P23919	558
CAD	UPRT	Q96BW1	529
CAD	CMPK1	P30085	517
CAD	PFAS	O15067	508
CAD	ADSS2	P30520	496
CAD	UCK1	Q9HA47	496

IntAct

235 interactions, top by confidence:

A	B	Type	Score
MAP3K14	CHUK	psi-mi:“MI:0914”(association)	0.950
ATG13	ULK1	psi-mi:“MI:2364”(proximity)	0.940
CFTR	ESYT2	psi-mi:“MI:0914”(association)	0.710
CFTR	ESYT2	psi-mi:“MI:2364”(proximity)	0.710
PKN3	ARHGAP10	psi-mi:“MI:0914”(association)	0.680
MAPK7	PFDN6	psi-mi:“MI:0914”(association)	0.640
NCBP2	KPNA3	psi-mi:“MI:0914”(association)	0.640
PPP5C	IRS4	psi-mi:“MI:0914”(association)	0.570
PPP5C	IRS4	psi-mi:“MI:2364”(proximity)	0.570
CAD	CAD	psi-mi:“MI:0407”(direct interaction)	0.560
ILK	HAX1	psi-mi:“MI:0914”(association)	0.530
TUBB3	POTEF	psi-mi:“MI:0914”(association)	0.530
GRB2	ARHGEF35	psi-mi:“MI:0914”(association)	0.530
PRKCZ	IPO5	psi-mi:“MI:0914”(association)	0.530
ILK	ILVBL	psi-mi:“MI:0914”(association)	0.530
ARRDC4	WWP2	psi-mi:“MI:0914”(association)	0.530
YWHAZ	BLTP3B	psi-mi:“MI:0914”(association)	0.530
CFTR	PLEKHG3	psi-mi:“MI:0914”(association)	0.480

BioGRID (542): CAD (Affinity Capture-MS), CAD (Affinity Capture-MS), CAD (Affinity Capture-MS), CAD (Reconstituted Complex), CAD (Affinity Capture-MS), CAD (Affinity Capture-MS), ACTR1A (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation), CAD (Co-fractionation)

ESM2 similar proteins: A0A168WVR6, A0A2Z4HPY4, A0L9N3, A1UR54, A1VWK8, A8GG79, B0KTG8, B0RTS9, B0Y565, B1YR44, B2FL74, B2RQC6, B2U9U9, B4EAW9, B4SRZ0, C1DD43, K0E4E5, P05990, P08955, P14300, P27708, P31961, P45414, P55610, P58669, P59966, P9WEH1, P9WMR2, P9WMR3, Q21VT1, Q2L0B4, Q2P3M7, Q30ZN1, Q3AQA7, Q3BUK3, Q3JRQ3, Q4UTW8, Q5H0N8, Q5UQA6, Q62JW4

Diamond homologs: A1CA18, A1CZ92, A2R9B9, A4G0Y3, A4SXM1, A4YI77, B2RQC6, B2URJ0, B4S5S1, B8GNX4, O27079, O28995, O60060, O66727, O93937, P05990, P07258, P07259, P07756, P08955, P0DA12, P0DA13, P14845, P20054, P22572, P27708, P31327, P36838, P58894, P63729, P63733, P63734, P63735, P77885, P87183, Q09794, Q0U5H7, Q18990, Q1DUF5, Q2H132

SIGNOR signaling

25 interactions.

A	Effect	B	Mechanism
MAPK1	up-regulates	CAD	phosphorylation
MAPK3	up-regulates	CAD	phosphorylation
PRKACA	down-regulates	CAD	phosphorylation
RPS6KB1	up-regulates	CAD	phosphorylation
CAD	“up-regulates quantity”	“carbamoyl phosphate(2-)”	“chemical modification”
CAD	“up-regulates activity”	“Pyrimidine biosynthesis”
RPS6K	“up-regulates activity”	CAD	phosphorylation
CAD	“down-regulates quantity”	glutamine	“chemical modification”
CAD	“down-regulates quantity”	“L-glutamine zwitterion”	“chemical modification”
CAD	“down-regulates quantity”	hydrogencarbonate	“chemical modification”
CAD	“up-regulates quantity”	(S)-dihydroorotate	“chemical modification”
CAD	“up-regulates quantity”	L-glutamate(1-)	“chemical modification”
CAD	“down-regulates quantity”	L-aspartate(1-)	“chemical modification”
CAD	“up-regulates quantity”	N-carbamoyl-L-aspartate(2-)	“chemical modification”
CAD	“down-regulates quantity”	N-carbamoyl-L-aspartate(2-)	“chemical modification”
RPS6KB1	“up-regulates activity”	CAD	phosphorylation
CAD	“down-regulates quantity”	“carbamoyl phosphate(2-)”	“chemical modification”
Gbeta	up-regulates	CAD	phosphorylation
PRKACA	unknown	CAD	phosphorylation
PRKACA	“down-regulates activity”	CAD	phosphorylation
CAD	“up-regulates activity”	CAD	phosphorylation
PPP1CA	“down-regulates activity”	CAD	dephosphorylation
PP1	“down-regulates activity”	CAD	dephosphorylation
ERK1/2	“up-regulates activity”	CAD	phosphorylation
PKA	“down-regulates activity”	CAD	phosphorylation

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 224 IntAct physical interaction partners (hypergeometric vs the genome-wide background, BH-FDR, gene-set size 15–500, ranked by fold). A functional readout of the neighbourhood — distinct from this gene’s own memberships above, and biased toward well-studied / hub proteins, so read it as themes rather than proof.

Reactome pathways:

Pathway	Partners	Fold	FDR
RIP-mediated NFkB activation via ZBP1	6	24.0×	4e-06
TRAF6 mediated NF-kB activation	7	19.0×	2e-06
TNF signaling	7	17.6×	4e-06
MAP kinase activation	9	16.5×	1e-07
TNFR1-induced NF-kappa-B signaling pathway	8	16.0×	1e-06
JNK (c-Jun kinases) phosphorylation and activation mediated by activated human TAK1	5	15.4×	3e-04
Toll Like Receptor 10 (TLR10) Cascade	12	15.4×	8e-10
Toll Like Receptor 5 (TLR5) Cascade	12	15.4×	8e-10

GO biological processes:

GO term	Partners	Fold	FDR
non-canonical NF-kappaB signal transduction	8	33.4×	2e-08
Fc-gamma receptor signaling pathway involved in phagocytosis	7	24.3×	3e-06
cytoplasmic pattern recognition receptor signaling pathway	5	21.9×	3e-04
canonical NF-kappaB signal transduction	11	19.9×	9e-09
tumor necrosis factor-mediated signaling pathway	9	14.7×	3e-06
negative regulation of protein-containing complex assembly	5	11.3×	6e-03
mitophagy	7	11.0×	3e-04
positive regulation of type I interferon production	5	10.4×	7e-03

Disease & clinical

Clinical variants and AI predictions

ClinVar

2342 variants total. Per-class counts are floors (≥ shown; pagination cap):

Classification	Count (floor)
Pathogenic	62
Likely pathogenic	32
Uncertain significance	723
Likely benign	1379
Benign	65

Top pathogenic / likely-pathogenic (30)

Variant ID	HGVS	Classification
1342296	NM_004341.5(CAD):c.2995G>A (p.Val999Met)	Pathogenic
1351531	NM_004341.5(CAD):c.135C>A (p.Tyr45Ter)	Pathogenic
1363933	NM_004341.5(CAD):c.5420dup (p.Asp1808fs)	Pathogenic
1439500	NM_004341.5(CAD):c.3088C>T (p.Gln1030Ter)	Pathogenic
1446574	NM_004341.5(CAD):c.1172del (p.Pro391fs)	Pathogenic
1453934	NM_004341.5(CAD):c.473del (p.Leu158fs)	Pathogenic
1454035	NM_004341.5(CAD):c.4246C>T (p.Arg1416Ter)	Pathogenic
1456472	NM_004341.5(CAD):c.6079C>T (p.Gln2027Ter)	Pathogenic
1457870	NM_004341.5(CAD):c.2174dup (p.Thr726fs)	Pathogenic
1908255	NM_004341.5(CAD):c.736C>T (p.Arg246Ter)	Pathogenic
1968982	NM_004341.5(CAD):c.4365_4368del (p.Asp1454_Cys1455insTer)	Pathogenic
2002049	NM_004341.5(CAD):c.1948C>T (p.Gln650Ter)	Pathogenic
2007350	NM_004341.5(CAD):c.1556_1565del (p.Ala519fs)	Pathogenic
203465	NM_004341.5(CAD):c.1843-1G>A	Pathogenic
2048288	NM_004341.5(CAD):c.2515C>T (p.Arg839Ter)	Pathogenic
2098540	NM_004341.5(CAD):c.1769del (p.Val590fs)	Pathogenic
2116615	NM_004341.5(CAD):c.2453G>A (p.Trp818Ter)	Pathogenic
2118936	NM_004341.5(CAD):c.2634del (p.Ala879fs)	Pathogenic
2153394	NM_004341.5(CAD):c.2169_2170del (p.Gly725fs)	Pathogenic
2694304	NM_004341.5(CAD):c.1418dup (p.Leu473fs)	Pathogenic
2695045	NM_004341.5(CAD):c.3976del (p.Ser1326fs)	Pathogenic
2698700	NM_004341.5(CAD):c.4324C>T (p.Gln1442Ter)	Pathogenic
2706674	NM_004341.5(CAD):c.4214_4220del (p.Leu1405fs)	Pathogenic
2712563	NM_004341.5(CAD):c.1330C>T (p.Gln444Ter)	Pathogenic
2748307	NM_004341.5(CAD):c.2210del (p.Val737fs)	Pathogenic
2749067	NM_004341.5(CAD):c.6387del (p.Phe2129fs)	Pathogenic
2758680	NM_004341.5(CAD):c.1435C>T (p.Gln479Ter)	Pathogenic
2759556	NM_004341.5(CAD):c.5599G>T (p.Glu1867Ter)	Pathogenic
2766717	NM_004341.5(CAD):c.5619_5620del (p.Arg1874fs)	Pathogenic
2798008	NM_004341.5(CAD):c.1543C>T (p.Arg515Ter)	Pathogenic

SpliceAI

6120 predictions. Top by Δscore:

Variant	Effect	Δscore
2:27217627:G:GT	donor_gain	1.0000
2:27217629:AGTGG:A	donor_loss	1.0000
2:27217630:GTGG:G	donor_gain	1.0000
2:27217632:GG:G	donor_gain	1.0000
2:27217632:GGGTA:G	donor_loss	1.0000
2:27217633:GG:G	donor_gain	1.0000
2:27217634:G:GA	donor_loss	1.0000
2:27217634:G:GG	donor_gain	1.0000
2:27217635:T:G	donor_loss	1.0000
2:27217996:GA:G	donor_gain	1.0000
2:27217997:A:G	donor_gain	1.0000
2:27218012:GCAAG:G	donor_loss	1.0000
2:27218013:CAAGG:C	donor_loss	1.0000
2:27218014:AAGG:A	donor_loss	1.0000
2:27218015:AG:A	donor_loss	1.0000
2:27218016:GG:G	donor_loss	1.0000
2:27218017:GTA:G	donor_loss	1.0000
2:27221214:ACAGT:A	acceptor_gain	1.0000
2:27221216:A:AG	acceptor_gain	1.0000
2:27221216:AGT:A	acceptor_gain	1.0000
2:27221217:G:GG	acceptor_gain	1.0000
2:27221217:GT:G	acceptor_gain	1.0000
2:27221217:GTG:G	acceptor_gain	1.0000
2:27222193:GGA:G	acceptor_gain	1.0000
2:27222657:CAAGG:C	donor_loss	1.0000
2:27222658:AAGG:A	donor_loss	1.0000
2:27222661:G:GG	donor_gain	1.0000
2:27222662:T:A	donor_loss	1.0000
2:27223915:A:AG	acceptor_gain	1.0000
2:27223916:G:GG	acceptor_gain	1.0000

AlphaMissense

14390 scored. Top likely-pathogenic:

Variant	Protein change	am_pathogenicity
2:27224470:T:C	F412L	1.000
2:27224472:T:A	F412L	1.000
2:27224472:T:G	F412L	1.000
2:27226902:T:A	W743R	1.000
2:27226902:T:C	W743R	1.000
2:27232644:A:C	S948R	1.000
2:27232646:C:A	S948R	1.000
2:27232646:C:G	S948R	1.000
2:27232653:T:C	F951L	1.000
2:27232655:T:A	F951L	1.000
2:27232655:T:G	F951L	1.000
2:27232657:A:T	D952V	1.000
2:27233087:A:C	S980R	1.000
2:27233089:C:A	S980R	1.000
2:27233089:C:G	S980R	1.000
2:27233513:T:A	W1065R	1.000
2:27233513:T:C	W1065R	1.000
2:27234216:T:C	L1203P	1.000
2:27235308:A:C	S1284R	1.000
2:27235310:T:A	S1284R	1.000
2:27235310:T:G	S1284R	1.000
2:27235541:A:C	K1325N	1.000
2:27235541:A:T	K1325N	1.000
2:27241944:A:C	S1973R	1.000
2:27241946:C:A	S1973R	1.000
2:27241946:C:G	S1973R	1.000
2:27224473:G:C	D413H	0.999
2:27224474:A:T	D413V	0.999
2:27224785:T:C	L432P	0.999
2:27224792:C:A	N434K	0.999

dbSNP variants (sampled 300 via entrez): RS1000181171 (2:27244202 A>G), RS1000250250 (2:27226790 T>C), RS1000251494 (2:27237655 G>A,T), RS1000301235 (2:27227237 C>A,G), RS1000356117 (2:27220174 T>C), RS1000559208 (2:27243081 C>G,T), RS1000636867 (2:27225592 G>A,C), RS1000747345 (2:27219392 A>T), RS1000856923 (2:27235886 A>C), RS1000884022 (2:27242465 A>G), RS1000929961 (2:27237299 C>G,T), RS1001095181 (2:27230253 G>A), RS1001096333 (2:27219110 C>G), RS1001417661 (2:27229746 C>G,T), RS1001486097 (2:27230714 A>C,G,T)

Disease associations

OMIM: gene MIM:114010 | disease phenotypes: MIM:616457, MIM:156000, MIM:610947

GenCC curated gene-disease

Disease	Classification	Inheritance
developmental and epileptic encephalopathy, 50	Definitive	Autosomal recessive

ClinGen Gene-Disease Validity (1)

Expert-panel classifications — Definitive > Strong > Moderate > Limited > Disputed > Refuted.

Disease	Classification	Inheritance
developmental and epileptic encephalopathy, 50	Definitive	AR

Mondo (6): developmental and epileptic encephalopathy, 50 (MONDO:0014647), infantile epileptic-dyskinetic encephalopathy (MONDO:0018226), Meniere disease (MONDO:0007972), coronary artery disease, autosomal dominant 2 (MONDO:0012586), intellectual disability (MONDO:0001071), epilepsy (MONDO:0005027)

Orphanet (4): CAD-CDG (Orphanet:448010), Infantile epileptic-dyskinetic encephalopathy (Orphanet:364063), NON RARE IN EUROPE: Menière disease (Orphanet:45360), NON RARE IN EUROPE: Unexplained intellectual disability (Orphanet:319658)

HPO phenotypes

25 total (25 of 25 shown, HPO-id order):

HPO	Term
HP:0000007	Autosomal recessive inheritance
HP:0001250	Seizure
HP:0001252	Hypotonia
HP:0001263	Global developmental delay
HP:0001508	Failure to thrive
HP:0001903	Anemia
HP:0001927	Acanthocytosis
HP:0001947	Renal tubular acidosis
HP:0001981	Schistocytosis
HP:0001987	Hyperammonemia
HP:0002014	Diarrhea
HP:0002015	Dysphagia
HP:0002133	Status epilepticus
HP:0002136	Broad-based gait
HP:0002283	Global brain atrophy
HP:0002376	Developmental regression
HP:0002465	Poor speech
HP:0003218	Oroticaciduria
HP:0003593	Infantile onset
HP:0003676	Progressive
HP:0003819	Death in childhood
HP:0004823	Anisopoikilocytosis
HP:0011463	Childhood onset
HP:0012345	Abnormal glycosylation
HP:0200134	Epileptic encephalopathy

GWAS associations

1 associations (top):

Study	Trait	p-value
GCST010244_62	Triglyceride levels	7.000000e-32

EFO canonical traits (1, from GWAS)

EFO ID	Trait name
EFO:0004530	triglyceride measurement

MeSH disease descriptors (5)

Descriptor	Name	Tree numbers
D004827	Epilepsy	C10.228.140.490
D008607	Intellectual Disability	C10.597.606.360; C23.888.592.604.646; F01.700.687; F03.625.539
D008575	Meniere Disease	C09.218.568.217.500
C567045	Coronary Artery Disease, Autosomal Dominant 2 (supp.)
C567924	Infantile Epileptic-Dyskinetic Encephalopathy (supp.)

Drugs & pharmacology

Drug and pharmacology data

Is drug target: yes

ChEMBL targets (1): CHEMBL3093 (SINGLE PROTEIN)

PharmGKB: 1 entry (VIP=true, CPIC=false)

ChEMBL bioactivities

10 potent at pChembl≥5 of 10 total, top 10 by pChembl (potency: 10 = 0.1 nM, 6 = 1 µM).

pChembl	Type	Value	Unit	Molecule
8.58	Kd	2.645	nM	CHEMBL3752910
8.58	ED50	2.645	nM	CHEMBL3752910
8.35	Kd	4.477	nM	CHEMBL5653589
8.35	ED50	4.477	nM	CHEMBL5653589
6.85	Ki	140	nM	CHEMBL29908
6.85	Ki	140	nM	CHEMBL170214
5.54	Ki	2900	nM	CHEMBL141678
5.51	Ki	3100	nM	CHEMBL343069
5.40	Ki	4000	nM	CHEMBL336479
5.29	Ki	5070	nM	CHEMBL168878

PubChem BioAssay actives

8 with measured affinity, of 21 total; 7 most potent distinct compounds. Largely complementary to BindingDB; screening values are coarse (µM, 4 dp), so sub-nM hits tie at the floor.

Compound	Assay	Type	Value	Unit
4-methyl-3-[(1-methyl-6-pyridin-3-ylpyrazolo[3,4-d]pyrimidin-4-yl)amino]-N-[3-(trifluoromethyl)phenyl]benzamide	2147979: Binding affinity to human CAD incubated for 45 mins by Kinobead based pull down assay	kd	0.0026	uM
4-methyl-3-[(2-methyl-6-pyridin-3-ylpyrazolo[3,4-d]pyrimidin-4-yl)amino]-N-[3-(trifluoromethyl)phenyl]benzamide	2147979: Binding affinity to human CAD incubated for 45 mins by Kinobead based pull down assay	kd	0.0045	uM
(4S,6R)-2-oxo-6-(sulfanylmethyl)-1,3-diazinane-4-carboxylic acid	55730: Compound ability to inhibit the catalytic activity of hamster Dihydroorotase (DHOase) expressed in Escherichia coli.	ki	0.1400	uM
2-oxo-4-(sulfanylmethyl)-5,6-dihydro-1H-pyrimidine-6-carboxylic acid	55732: Inhibitory activity against Dihydroorotase (DHO) at pH 7.4 and 8.5	ki	0.1400	uM
(2R)-4-hydroxy-6-oxo-4-sulfanylidene-1,4lambda5-azaphosphinane-2-carboxylic acid	55730: Compound ability to inhibit the catalytic activity of hamster Dihydroorotase (DHOase) expressed in Escherichia coli.	ki	2.9000	uM
4-hydroxy-4,6-dioxo-1,4lambda5-azaphosphinane-2-carboxylic acid	55730: Compound ability to inhibit the catalytic activity of hamster Dihydroorotase (DHOase) expressed in Escherichia coli.	ki	4.0000	uM
[2-(carbamoylamino)-3-ethoxy-3-oxopropyl]boronic acid	55731: Inhibitory activity against Dihydroorotase (DHO) at pH 6.0	ki	5.0700	uM

CTD chemical–gene interactions

77 total (human), top 30 by PubMed support.

Chemical	Actions (top 5)	PubMed papers
sodium arsenite	affects expression, decreases expression, increases abundance, increases expression	5
Estradiol	affects binding, decreases reaction, increases reaction, decreases phosphorylation, increases expression	4
bisphenol A	decreases expression	3
bisphenol F	increases expression, affects cotreatment, decreases expression	2
(+)-JQ1 compound	decreases expression	2
Benzo(a)pyrene	increases expression, increases mutagenesis	2
aristolochic acid I	increases expression	1
GSK-J4	decreases expression	1
FR900359	decreases phosphorylation	1
dicrotophos	increases expression	1
2,4,6-tribromophenol	increases expression	1
triphenyl phosphate	affects expression	1
alpha-pinene	decreases expression, increases abundance, affects cotreatment	1
pyrogallol 1,3-dimethyl ether	affects cotreatment, affects localization, decreases expression, increases expression	1
decabromobiphenyl ether	increases expression	1
trichostatin A	decreases expression	1
beta-lapachone	decreases expression	1
mono-(2-ethylhexyl)phthalate	increases expression	1
cobaltous chloride	decreases expression	1
tetrabromobisphenol A	increases expression	1
manganese chloride	decreases expression, increases abundance	1
ochratoxin A	increases expression	1
potassium chromate(VI)	increases expression	1
methacrylaldehyde	affects cotreatment, decreases expression, increases abundance	1
di-n-butylphosphoric acid	affects expression	1
perfluorooctane sulfonic acid	increases expression	1
CGP 52608	affects binding, increases reaction	1
2-palmitoylglycerol	increases expression	1
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide	decreases expression, affects cotreatment	1
belinostat	decreases expression	1

ChEMBL screening assays

17 unique, capped per target: 16 binding, 1 admet

Representative assays (with source publication via chembl_document):

Assay ID	Type	Description	Source paper
CHEMBL4118734	Binding	Binding affinity to CAD in human NCI-H23 cells at 1 uM by mass spectrometry based pull down assay	Studies of TAK1-centered polypharmacology with novel covalent TAK1 inhibitors. — Bioorg Med Chem
CHEMBL4619225	ADMET	Inhibition of CAD (unknown origin) CSP2 activity expressed in Escherichia coli BL21 (DE3) preincubated for 20 mins followed by ATP addition measured after 60 mins by ADP-Glo assay	Discovery of 2,6-Dimethylpiperazines as Allosteric Inhibitors of CPS1. — ACS Med Chem Lett

Cellosaurus cell lines

5 cell lines: 4 cancer cell line, 1 induced pluripotent stem cell

First 10 cell lines (id-ordered, not curated):

Cellosaurus	Name	Category	Sex
CVCL_C4S7	BCHNEi001-A	Induced pluripotent stem cell	Male
CVCL_D9Z5	Ubigene HeLa CAD KO	Cancer cell line	Female
CVCL_XM42	HAP1 CAD (-)	Cancer cell line	Male
CVCL_ZD03	Mel 04.01	Cancer cell line	Male
CVCL_ZD06	Mel 12.07	Cancer cell line	Male

Clinical trials (associated diseases)

300 trials via MONDO — disease-level, not drug-specific.

Trial	Phase	Status	Title
NCT01574313	PHASE4	COMPLETED	Effect of Stellate Ganglion Block on Meniere’s Disease
NCT02529475	PHASE4	TERMINATED	Evaluation of Inner Ear and Brain Structures With Contrast-enhanced MRI in Healthy Subjects (HYDROPS)
NCT04815187	PHASE4	ACTIVE_NOT_RECRUITING	Repurposed Use of Allergic Rhinitis and Allergic Asthma Drug to Reduce Vertigo and Hearing Loss in Meniere’s Disease
NCT05657860	PHASE4	COMPLETED	Guanfacine Extended Release for the Reduction of Aggression and Self-injurious Behavior Associated With Prader-Willi Syndrome
NCT05744479	PHASE4	RECRUITING	Metformin for Antipsychotic-induced Weight Gain in Adults With Intellectual Disability
NCT06107829	PHASE4	WITHDRAWN	Valbenazine Treatment of Tardive Dyskinesia in Adults With Intellectual/Developmental Disabilities
NCT06997198	PHASE4	NOT_YET_RECRUITING	Deutetrabenazine Treatment for Tardive Dyskinesia in Intellectual/Developmental Disabilities
NCT00004637	PHASE4	COMPLETED	Double-Blind, Placebo-Controlled Trial of Vitamin E as Add-on Therapy for Children With Epilepsy
NCT00043914	PHASE4	COMPLETED	Measurement Of Serum Levels Of Two Antiepileptic Drugs During Conversion In Patients With Epilepsy
NCT00132223	PHASE4	UNKNOWN	Effects on the Diagnostic Accuracy of Magnetic Imaging Angiographies of the Supra-Aortic Vessels by Three Different Magnetic Resonance Contrast Agents in Patients
NCT00133081	PHASE4	UNKNOWN	Study to Improve the Treatment of Epilepsy (SITE)
NCT00137709	PHASE4	UNKNOWN	Hormone Profiles in Adults With Newly Diagnosed Epilepsy
NCT00154076	PHASE4	COMPLETED	A Multicenter Comparative Trial of Zonisamide and Topiramate as Initial Monotherapy in Untreated Epilepsies
NCT00165828	PHASE4	TERMINATED	Efficacy and Safety of an add-on Treatment With Zonisamide in Adults With Focal Epileptic Seizures With or Without Secondary Generalization
NCT00181116	PHASE4	COMPLETED	Levetiracetam for Benign Rolandic Epilepsy
NCT00207935	PHASE4	COMPLETED	Use of Sustained Release Antiepileptic Medication (Depakote® ER) for Pediatric Epilepsy in a Mental Retardation/Developmental Disorder Population
NCT00215592	PHASE4	COMPLETED	Open Label, Zonegran (Zonisamide) In Partial Onset Seizures
NCT00266604	PHASE4	COMPLETED	A Study to Evaluate the Dosing, Effectiveness and Safety of Topiramate for the Treatment of Epilepsy
NCT00288639	PHASE4	COMPLETED	Lyrica (Pregabalin) Administered as an Add-on Therapy for Partial Seizures (LEADER).
NCT00312676	PHASE4	UNKNOWN	Compare Tolerability of an Overnight Switch to Gradual Switch Between Two Different Forms of Depakote
NCT00323947	PHASE4	COMPLETED	Methylphenidate for Treating Attention Deficit Hyperactivity Disorder in Children With Both ADHD and Epilepsy
NCT00385411	PHASE4	COMPLETED	Study of Valproate in Young Patients Suffering From Epilepsy
NCT00522418	PHASE4	TERMINATED	Study Comparing Best Medical Practice With or Without VNS Therapy in Pharmacoresistant Partial Epilepsy Patients
NCT00537940	PHASE4	COMPLETED	Comparative Study Of Pregabalin And Gabapentin As Adjunctive Therapy In Subjects With Partial Seizures
NCT00552526	PHASE4	UNKNOWN	Ketogenic Diet vs.Antiepileptic Drug Treatment in Drug Resistant Epilepsy
NCT00564915	PHASE4	COMPLETED	RCT of the Efficacy of the Ketogenic Diet in the Treatment of Epilepsy
NCT00571155	PHASE4	COMPLETED	Trial of Levetiracetam in Patients With Primary Brain Tumors and Symptomatic Seizures Who Undergo Surgery
NCT00572195	PHASE4	COMPLETED	RNS® System LTT Study
NCT00610532	PHASE4	TERMINATED	Evaluating the Transporter Protein Inhibitor Probenecid In Patients With Epilepsy
NCT00630357	PHASE4	COMPLETED	Trial to Evaluate the Safety and Efficacy of Keppra After Conversion to Mono-therapy in Subjects With Partial Epilepsy
NCT00630630	PHASE4	COMPLETED	Study on Safety and Efficacy of Levetiracetam in the Adjunctive Treatment of Female Subjects With C1 Catamenial Epilepsy
NCT00630968	PHASE4	COMPLETED	S.K.A.T.E.: Safety of Keppra as Adjunctive Therapy in Epilepsy
NCT00631150	PHASE4	COMPLETED	A Phase IV-Pharmacovigilance Study of Keppra Greece - S.K.A.T.E.: Safety of Keppra as Adjunctive Therapy in Epilepsy
NCT00659958	PHASE4	COMPLETED	ZAGAL Study: Evaluating Effectiveness and Tolerability of Zonisamide as Adjunctive Therapy in Patients With Partial Onset Seizures Treated With Two Antiepileptic Drugs
NCT00713622	PHASE4	COMPLETED	Comparing The Effect On Cognition Of Adjunctive Therapy With Zonisamide Versus Sodium Valproate
NCT00807989	PHASE4	COMPLETED	The Efficacy and Safety of Low Dose Combination of LTG and VPA Compared to CBZ Monotherapy
NCT00832884	PHASE4	COMPLETED	The Safety of Intravenous Lacosamide
NCT00869622	PHASE4	COMPLETED	Antiepileptic Drugs and Osteoporotic Prevention Trial
NCT00896987	PHASE4	COMPLETED	Lamotrigine Cognitive Function Study in Adult Untreated Epilepsies
NCT00952081	PHASE4	COMPLETED	A Pilot Study to Evaluate Efficacy and Safety of Clevidipine in Neurosurgical Patients