FOLH1

Summary

FOLH1 (folate hydrolase 1, HGNC:3788) is a protein-coding gene on chromosome 11p11.12, encoding Glutamate carboxypeptidase 2 (Q04609). Has both folate hydrolase and N-acetylated-alpha-linked-acidic dipeptidase (NAALADase) activity.

This gene encodes a type II transmembrane glycoprotein belonging to the M28 peptidase family. The protein acts as a glutamate carboxypeptidase on different alternative substrates, including the nutrient folate and the neuropeptide N-acetyl-l-aspartyl-l-glutamate and is expressed in a number of tissues such as prostate, central and peripheral nervous system and kidney. A mutation in this gene may be associated with impaired intestinal absorption of dietary folates, resulting in low blood folate levels and consequent hyperhomocysteinemia. Expression of this protein in the brain may be involved in a number of pathological conditions associated with glutamate excitotoxicity. In the prostate the protein is up-regulated in cancerous cells and is used as an effective diagnostic and prognostic indicator of prostate cancer. This gene likely arose from a duplication event of a nearby chromosomal region. Alternative splicing gives rise to multiple transcript variants encoding several different isoforms.

Source: NCBI Gene 2346 — RefSeq curated summary.

At a glance

• GWAS associations: 22
• Clinical variants (ClinVar): 93 total
• Druggable target: yes — 7 molecules with ChEMBL bioactivity
• MANE Select transcript: NM_004476

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:3788
Approved symbol	FOLH1
Name	folate hydrolase 1
Location	11p11.12
Locus type	gene with protein product
Status	Approved
Aliases	PSM, PSMA, NAALAD1, NAALAdase, GCP2, GCPII
Ensembl gene	ENSG00000086205
Ensembl biotype	protein_coding
OMIM	600934
Entrez	2346

Gene structure

Transcript identifiers

Ensembl transcripts: 18 — 10 protein_coding, 4 retained_intron, 3 nonsense_mediated_decay, 1 protein_coding_CDS_not_defined

ENST00000256999, ENST00000340334, ENST00000356696, ENST00000458311, ENST00000525629, ENST00000525826, ENST00000526226, ENST00000529117, ENST00000529646, ENST00000529648, ENST00000532018, ENST00000533034, ENST00000533510, ENST00000897368, ENST00000897369, ENST00000897370, ENST00000953048, ENST00000953049

RefSeq mRNA: 6 — MANE Select: NM_004476 NM_001014986, NM_001193471, NM_001193472, NM_001193473, NM_001351236, NM_004476

CCDS: CCDS31493, CCDS53627, CCDS53628, CCDS7946

Canonical transcript exons

ENST00000256999 — 19 exons

Exon	Start	End
ENSE00001627795	49185669	49185855
ENSE00003459309	49156717	49156807
ENSE00003471104	49173357	49173476
ENSE00003471979	49171195	49171277
ENSE00003489243	49175859	49175957
ENSE00003510751	49169195	49169258
ENSE00003521929	49153846	49153927
ENSE00003539450	49200255	49200441
ENSE00003554421	49148639	49148731
ENSE00003555470	49164705	49164772
ENSE00003558288	49174892	49174977
ENSE00003590268	49206067	49206172
ENSE00003596455	49192793	49192894
ENSE00003639838	49186644	49186769
ENSE00003641088	49154228	49154492
ENSE00003656272	49208292	49208602
ENSE00003660926	49183149	49183242
ENSE00003678748	49157952	49158043
ENSE00003693627	49145092	49146945

Expression profiles

Bgee: expression breadth ubiquitous, 138 present calls, max score 99.33.

FANTOM5 (CAGE): breadth broad, TPM avg 5.2900 / max 482.8214, expressed in 278 samples.

FANTOM5 promoters (9 alternative TSS)

Promoter ID	TPM avg	Samples expressed
119687	1.2420	200
119686	1.0220	124
119684	0.9753	116
119683	0.9321	92
119689	0.5186	104
119688	0.2303	109
206281	0.1544	57
119685	0.1269	69
119682	0.0885	53

Top tissues by expression

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

Tissue	Anatomy ID	Expression score	Quality
duodenum	UBERON:0002114	99.33	gold quality
corpus callosum	UBERON:0002336	97.43	gold quality
C1 segment of cervical spinal cord	UBERON:0006469	96.45	gold quality
spinal cord	UBERON:0002240	95.92	gold quality
substantia nigra	UBERON:0002038	93.27	gold quality
Ammon’s horn	UBERON:0001954	91.31	gold quality
putamen	UBERON:0001874	90.67	gold quality
prostate gland	UBERON:0002367	89.90	gold quality
amygdala	UBERON:0001876	88.72	gold quality
temporal lobe	UBERON:0001871	88.27	gold quality
liver	UBERON:0002107	88.08	gold quality
gall bladder	UBERON:0002110	86.91	gold quality
caudate nucleus	UBERON:0001873	86.58	gold quality
ventricular zone	UBERON:0003053	85.96	gold quality
right lobe of liver	UBERON:0001114	85.60	gold quality
primary visual cortex	UBERON:0002436	85.21	gold quality
hypothalamus	UBERON:0001898	85.17	gold quality
Brodmann (1909) area 9	UBERON:0013540	84.93	gold quality
ganglionic eminence	UBERON:0004023	82.80	gold quality
prefrontal cortex	UBERON:0000451	82.56	gold quality
nucleus accumbens	UBERON:0001882	82.24	gold quality
male germ line stem cell (sensu Vertebrata) in testis	CL:0000089 ∩ UBERON:0000473	81.88	gold quality
cerebral cortex	UBERON:0000956	81.58	gold quality
saliva-secreting gland	UBERON:0001044	81.46	gold quality
frontal cortex	UBERON:0001870	81.07	gold quality
dorsolateral prefrontal cortex	UBERON:0009834	80.69	gold quality
right frontal lobe	UBERON:0002810	80.18	gold quality
minor salivary gland	UBERON:0001830	80.02	gold quality
adult mammalian kidney	UBERON:0000082	79.38	gold quality
central nervous system	UBERON:0001017	79.21	gold quality

Single-cell (SCXA)

Detected in 3 experiment(s), a significant marker in 3.

Experiment	Marker?	Max mean expression
E-ANND-3	yes	15.82
E-MTAB-6678	yes	12.80
E-GEOD-84465	yes	9.75

Regulation

Is transcription factor: no

Upstream regulators (CollecTRI, top): AR, ELF3, ELF5, ETS1, NFATC1, VDR

miRNA regulators (miRDB)

8 targeting FOLH1, 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-4282	99.99	75.36	6408
HSA-MIR-607	99.97	73.62	5593
HSA-MIR-3671	99.90	73.04	3897
HSA-MIR-140-3P	99.04	67.69	1324
HSA-MIR-4272	98.76	68.74	1810
HSA-MIR-660-5P	98.16	68.27	680
HSA-MIR-1236-5P	96.62	66.38	856
HSA-MIR-5591-3P	96.23	67.03	489

Literature-anchored findings (GeneRIF, showing 40)

• expressed in prostate tissue, and especially in prostate cancer tissue (PMID:12474535)
• GCP2 polymorphism explained nearly 50% of the variance of red blood cell folate in ESRD (PMID:12707400)
• GCP2 1561C>T is associated with elevated folate levels but not homocysteine levels in kidney transplant patients (PMID:12753319)
• PSMA expression is regulated by NFATc1 with an AP-3 binding site (PMID:12850144)
• N-linked carbohydrate structures are important for the folate hydrolase function of the protein. Removal of sugars partially or completely causes PSMA to be enzymatically inactive, improperly folded, resulting in increased rate of degradation. (PMID:12949938)
• PSMA is internalized via a clathrin-dependent endocytic mechanism and that internalization of PSMA is mediated by the five N-terminal amino acids (PMID:14528023)
• proteins binding PSMA (PMID:15141017)
• Analysis of the predicted N-glycosylation sites also provides evidence that these sites are critical for GCPII carboxypeptidase activity (PMID:15152093)
• amino acid segments at the N- and C-termini of the ectodomain of GCPII are essential for its carboxypeptidase activity and/or proper folding (PMID:15206943)
• PSMA gene lacks specificity for detecting prostate epithelial cells (PMID:15389976)
• Knockdown of PSMA expression increased invasiveness of LNCaP cells by 5-fold. (PMID:15705868)
• Transcriptional activity of the PSMA promoter/enhancer is prostate specific. (PMID:15713827)
• Structure and relevance to the development of chemotherapeutics and cancer-imaging agents. (PMID:15837926)
• PSMA may function as a receptor internalizing a putative ligand, an enzyme playing a role in nutrient uptake, and a peptidase involved in signal transduction in prostate epithelial cells. [review] (PMID:15840561)
• acquisition of folding determinants in the Golgi is an essential prerequisite for protein trafficking and sorting of PSMA (PMID:16221666)
• The folate hydrolase activity of PSMA may provide a growth advantage to prostate cancer cells in low folate and physiological folate environments. (PMID:16496414)
• PSMA binds to caveolin-1 and undergoes internalization via a caveolae-dependent mechanism in microvascular endothelial cells (PMID:16713605)
• In suicide gene therapy, anti-PSMA-liposome complex exerted a significant inhibitory effect on the growth of LNCaP xenograft, in contrast to normal IgG-liposome complex. (PMID:17032155)
• Using pure recombinant GCPII and homologous GCPIII, we conclude that GCPII is responsible for the majority of overall NAAG-hydrolyzing activity in the human brain. (PMID:17150306)
• Polymorphism is associated with higher plasma folate and lower homocysteine concentrations in women who abstain from alcohol. (PMID:17684227)
• Pattern differences in GCP II gene promoter expression in SVG and LNCaP cells suggest that sequences beyond 240 bp may be important for tissue-specific GCP II expression. (PMID:17689503)
• the residues forming the S1 pocket might be more important for the ‘fine-tuning’ of GCPII substrate specificity (PMID:17714508)
• the intracellular transport of PSMA occurs through populations of DRMs distinct for each biosynthetic form and cellular compartment (PMID:17935484)
• Tissues such as prostate and testes exhibit different GCPII expression levels among the species studied (human, rats, swine). (PMID:18076021)
• Crystal structures of human GCPII in complex with phosphapeptide analogs show the S1 pocket of GCPII and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. (PMID:18234225)
• prostate-specific membrane antigen is expressed in tumor-associated neovasculature of the majority of renal cortical tumors (PMID:18344976)
• PSMA is underexpressed in advanced stage endometrial adenocarcinoma (EAC); loss of PSMA expression can be considered as a prognostic marker in patients with EAC; loss of PSMA expression in a subset of EAC cases could be due to epigenetic silencing (PMID:18349274)
• Given our finding of frequent expression of PSMA in Schwannomas, they should be clinically considered in the differential diagnosis of a lesion that is positive on PSMA radioimmunoscintigraphy study performed during a metastatic work-up of PCa patient. (PMID:18534872)
• the efficacy of 5-FU-based chemotherapy in prostate cancers can be significantly improved by targeted expression of the suicide gene UPRT under the control of prostate-specific membrane antigen promoter/enhancer (PMID:18626508)
• The present study describes an efficient RNAi system for gene silencing that is specific to prostate cancer cells using the PSMA promoter/enhancer (PMID:18639471)
• Neonatal methylmalonic acid was predicted by maternal methylmalonic acid and GCPII (PMID:18823966)
• prostate-specific membrane antigen is expressed in neovasculature from physiologic regenerative and reparative conditions (PMID:18839017)
• PSMA was the most useful marker to identify residual prostatic carcinoma after hormone therapy. (PMID:18844933)
• Ectopic PSMA expression on PC-3 cells increased the invasive capacity of cells in in vitro invasion assays, which could be competed out by folic acid. These results suggest PSMA facilitates the development of prostate cancer (PMID:18974153)
• analysis of interactions between human glutamate carboxypeptidase II and urea-based inhibitors (PMID:19053759)
• PSM’ protein is likely not generated by alternative splicing of the PSMA gene but by different mechanism, probably via an endoproteolytic cleavage of the full-length PSMA. (PMID:19107881)
• although the RFC1 80G > A and FOLH1 1561C > T polymorphisms may influence folate status, they are not likely to have a major independent role in the development of colorectal cancer (PMID:19172696)
• PSMA-fostered RAS-RAC1-MAPK pathway activation produced a strong induction of NF-kappaB activation associated with an increased expression of IL-6 and CCL5 genes. (PMID:19242540)
• Reaction mechanisms of GCPII revealed by mutagenesis, X-ray crystallography, and computational methods. (PMID:19301871)
• PSMA and prostate stem cell antigen are both highly expressed in lymph node and bone metastases of prostate cancer. (PMID:19343734)

Cross-species orthologs

5 orthologs

Organism	Symbol	Gene ID
danio_rerio	naalad2	ENSDARG00000021383
mus_musculus	Folh1	ENSMUSG00000001773
rattus_norvegicus	Folh1	ENSRNOG00000013770
caenorhabditis_elegans		WBGENE00007954
caenorhabditis_elegans		WBGENE00020082

Paralogs (5): TFRC (ENSG00000072274), NAALAD2 (ENSG00000077616), TFR2 (ENSG00000106327), NAALADL1 (ENSG00000168060), NAALADL2 (ENSG00000177694)

Protein

Protein identifiers

Glutamate carboxypeptidase 2 — Q04609 (reviewed: Q04609)

Alternative names: Cell growth-inhibiting gene 27 protein, Folate hydrolase 1, Folylpoly-gamma-glutamate carboxypeptidase, Glutamate carboxypeptidase II, Membrane glutamate carboxypeptidase, N-acetylated-alpha-linked acidic dipeptidase I, Prostate-specific membrane antigen, Pteroylpoly-gamma-glutamate carboxypeptidase

All UniProt accessions (5): Q04609, E9PI29, E9PKM3, E9PLV0, E9PMK6

UniProt curated annotations — full annotation on UniProt →

Function. Has both folate hydrolase and N-acetylated-alpha-linked-acidic dipeptidase (NAALADase) activity. Has a preference for tri-alpha-glutamate peptides. In the intestine, required for the uptake of folate. In the brain, modulates excitatory neurotransmission through the hydrolysis of the neuropeptide, N-aceylaspartylglutamate (NAAG), thereby releasing glutamate. Involved in prostate tumor progression. Also exhibits a dipeptidyl-peptidase IV type activity. In vitro, cleaves Gly-Pro-AMC.

Subunit / interactions. Homodimer.

Subcellular location. Cell membrane Cytoplasm.

Tissue specificity. Highly expressed in prostate epithelium. Detected in urinary bladder, kidney, testis, ovary, fallopian tube, breast, adrenal gland, liver, esophagus, stomach, small intestine, colon and brain (at protein level). Detected in the small intestine, brain, kidney, liver, spleen, colon, trachea, spinal cord and the capillary endothelium of a variety of tumors. Expressed specifically in jejunum brush border membranes. In the brain, highly expressed in the ventral striatum and brain stem. Also expressed in fetal liver and kidney. Isoform PSMA’ is the most abundant form in normal prostate. Isoform PSMA-1 is the most abundant form in primary prostate tumors. Isoform PSMA-9 is specifically expressed in prostate cancer.

Post-translational modifications. The first two amino acids at the N-terminus of isoform PSMA’ appear to be cleaved by limited proteolysis. The N-terminus is blocked.

Activity regulation. The NAALADase activity is inhibited by beta-NAAG, quisqualic acid, 2-(phosphonomethyl) pentanedioic acid (PMPA) and EDTA. Activated by cobalt.

Cofactor. Binds 2 Zn(2+) ions per subunit. Required for NAALADase activity.

Domain organisation. The NAALADase activity is found in the central region, the dipeptidyl peptidase IV type activity in the C-terminal.

Induction. In the prostate, up-regulated in response to androgen deprivation.

Polymorphism. Genetic variation in FOLH1 may be associated with low folate levels and consequent hyperhomocysteinemia. This condition can result in increased risk of cardiovascular disease, neural tube defects, and cognitive deficits.

Miscellaneous. PSMA is used as a diagnostic and prognostic indicator of prostate cancer, and as a possible marker for various neurological disorders such as schizophrenia, Alzheimer disease and Huntington disease.

Similarity. Belongs to the peptidase M28 family. M28B subfamily.

Isoforms (8)

UniProt ID	Names	Canonical?
Q04609-1	PSMA-1	yes
Q04609-3	PSMA-3
Q04609-4	PSMA-4
Q04609-6	PSMA’
Q04609-7	PSMA-7
Q04609-8	PSMA-8
Q04609-9	PSMA-9, PSM-E
Q04609-10	10

RefSeq proteins (6): NP_001014986, NP_001180400, NP_001180401, NP_001180402, NP_001338165, NP_004467* (*=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR003137	PA_domain	Domain
IPR007365	TFR-like_dimer_dom	Domain
IPR007484	Peptidase_M28	Domain
IPR036757	TFR-like_dimer_dom_sf	Homologous_superfamily
IPR039373	Peptidase_M28B	Family
IPR046450	PA_dom_sf	Homologous_superfamily

Pfam: PF02225, PF04253, PF04389

Enzyme classification (BRENDA):

• EC 3.4.17.21 — glutamate carboxypeptidase II (BRENDA: 10 organisms, 60 substrates, 336 inhibitors, 15 Km, 14 kcat entries)

Substrate kinetics (BRENDA)

5 substrates with measured Km, best-characterized 5. Km ranges are aggregated across organisms/conditions.

Substrate	Km (mM)	Measurements
N-ACETYL-L-ASP-L-GLU	0.0001–540	6
N-ACETYL-ALPHA-L-ASPARTYL-L-GLUTAMATE	0.0001–0.0003	4
N-ACETYL-L-ASPARTYL-L-GLUTAMATE	0.0006–0.0019	2
PTEROYL-DI-L-GLUTAMATE	—	2
N-ACETYL-ASP-GLU	130	1

UniProt features (140 total): strand 30, helix 28, mutagenesis site 23, binding site 19, glycosylation site 10, splice variant 7, turn 6, sequence variant 4, active site 4, sequence conflict 3, topological domain 2, chain 1, transmembrane region 1, modified residue 1, region of interest 1

Structure

Experimental structures (PDB)

88 structures, top 30 by resolution.

PDB	Method	Resolution (Å)
5O5T	X-RAY DIFFRACTION	1.43
5OF0	X-RAY DIFFRACTION	1.48
3BI1	X-RAY DIFFRACTION	1.5
6FE5	X-RAY DIFFRACTION	1.52
5O5U	X-RAY DIFFRACTION	1.53
3D7F	X-RAY DIFFRACTION	1.54
3D7H	X-RAY DIFFRACTION	1.55
8BO8	X-RAY DIFFRACTION	1.55
8BOL	X-RAY DIFFRACTION	1.55
6SGP	X-RAY DIFFRACTION	1.58
6SKH	X-RAY DIFFRACTION	1.58
8BOW	X-RAY DIFFRACTION	1.58
2XEF	X-RAY DIFFRACTION	1.59
2XEG	X-RAY DIFFRACTION	1.59
3BHX	X-RAY DIFFRACTION	1.6
3RBU	X-RAY DIFFRACTION	1.6
6EZ9	X-RAY DIFFRACTION	1.61
2OR4	X-RAY DIFFRACTION	1.62
4NGP	X-RAY DIFFRACTION	1.63
6F5L	X-RAY DIFFRACTION	1.63
2OOT	X-RAY DIFFRACTION	1.64
4NGN	X-RAY DIFFRACTION	1.64
4W9Y	X-RAY DIFFRACTION	1.64
3SJF	X-RAY DIFFRACTION	1.65
3SJG	X-RAY DIFFRACTION	1.65
4MCP	X-RAY DIFFRACTION	1.65
4MCR	X-RAY DIFFRACTION	1.65
4OC2	X-RAY DIFFRACTION	1.65
4P4D	X-RAY DIFFRACTION	1.65
5O5R	X-RAY DIFFRACTION	1.65

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-Q04609-F1	93.89	0.89

Antibody-complex structures (SAbDab): 3 — 9HLW, 9HVI, 9HVK

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): 424 (nucleophile; for naaladase activity); 628 (charge relay system); 666 (charge relay system); 689 (charge relay system)

Ligand- & substrate-binding residues (19): 257; 269; 272; 377; 387; 387; 424; 425; 433; 436; 453; 517–518 …

Post-translational modifications (1): 10

Glycosylation sites (10): 51, 76, 121, 140, 153, 195, 336, 459, 476, 638

Mutagenesis-validated functional residues (23):

Position	Phenotype
51	loss of glycosylation. reduces enzyme activity.
76	loss of glycosylation. reduces enzyme activity.
121	loss of glycosylation. severely reduced enzyme activity.
140	loss of glycosylation. severely reduced enzyme activity.
153	loss of glycosylation. severely reduced enzyme activity.
195	loss of glycosylation. severely reduced enzyme activity.
336	loss of glycosylation. reduces enzyme activity.
377	complete loss of activity.
379	complete loss of activity.
387	complete loss of activity.
387	no effect on enzyme activity.
388	no effect on enzyme activity.
424	complete loss of activity.
424	reduces enzyme activity.
425	complete loss of activity.
453	complete loss of activity.
453	reduces enzyme activity.
454	reduces enzyme activity.
459	loss of glycosylation. reduces enzyme activity.
476	loss of glycosylation. reduces enzyme activity.
638	loss of glycosylation. abolishes enzyme activity.
640	abolishes enzyme activity.

Function

Pathways and Gene Ontology

Reactome pathways

1 pathways

ID	Pathway
R-HSA-8963693	Aspartate and asparagine metabolism

MSigDB gene sets: 96 (showing top): GSE18804_BRAIN_VS_COLON_TUMORAL_MACROPHAGE_DN, MODULE_172, GOBP_DIGESTION, GOBP_GLUTAMINE_FAMILY_AMINO_ACID_BIOSYNTHETIC_PROCESS, GOMF_METALLOPEPTIDASE_ACTIVITY, GOBP_ALPHA_AMINO_ACID_METABOLIC_PROCESS, GOBP_GLUTAMATE_METABOLIC_PROCESS, ASTON_MAJOR_DEPRESSIVE_DISORDER_DN, ROVERSI_GLIOMA_COPY_NUMBER_UP, GOCC_CELL_SURFACE, GOBP_AMINO_ACID_BIOSYNTHETIC_PROCESS, GOBP_GLUTAMINE_FAMILY_AMINO_ACID_METABOLIC_PROCESS, GOBP_ORGANIC_ACID_BIOSYNTHETIC_PROCESS, RODWELL_AGING_KIDNEY_NO_BLOOD_DN, GOBP_DICARBOXYLIC_ACID_METABOLIC_PROCESS

GO Biological Process (5): proteolysis (GO:0006508), obsolete glutamate biosynthetic process (GO:0006537), folic acid-containing compound metabolic process (GO:0006760), intestinal folate absorption (GO:0098829), positive regulation of glutamate receptor signaling pathway (GO:1900451)

GO Molecular Function (11): carboxypeptidase activity (GO:0004180), metallocarboxypeptidase activity (GO:0004181), peptidase activity (GO:0008233), dipeptidase activity (GO:0016805), metal ion binding (GO:0046872), Ac-Asp-Glu binding (GO:1904492), tetrahydrofolyl-poly(glutamate) polymer binding (GO:1904493), catalytic activity (GO:0003824), protein binding (GO:0005515), metallopeptidase activity (GO:0008237), hydrolase activity (GO:0016787)

GO Cellular Component (5): cytoplasm (GO:0005737), plasma membrane (GO:0005886), cell surface (GO:0009986), membrane (GO:0016020), extracellular exosome (GO:0070062)

Reactome top-level categories

Rollup of top-1 pathways:

Category	Pathways
Metabolism of amino acids and derivatives	1

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
cellular anatomical structure	3
exopeptidase activity	2
protein metabolic process	1
modified amino acid metabolic process	1
pteridine-containing compound metabolic process	1
intestinal absorption	1
glutamate receptor signaling pathway	1
positive regulation of signal transduction	1
regulation of glutamate receptor signaling pathway	1
carboxypeptidase activity	1
metalloexopeptidase activity	1
hydrolase activity	1
catalytic activity, acting on a protein	1
cation binding	1
anion binding	1
oligopeptide binding	1
small molecule binding	1
molecular_function	1
binding	1
peptidase activity	1
catalytic activity	1
intracellular anatomical structure	1
membrane	1
cell periphery	1
extracellular vesicle	1

Protein interactions and networks

STRING

1786 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
FOLH1	KLK3	P07288	939
FOLH1	PSCA	O43653	910
FOLH1	ERBB2	P04626	824
FOLH1	GRPR	P30550	819
FOLH1	FOLR1	P15328	817
FOLH1	FOLR3	P41439	798
FOLH1	ALB	P02768	797
FOLH1	IZUMO1R	A6ND01	790
FOLH1	FOLR2	P14207	783
FOLH1	EGFR	P00533	780
FOLH1	GRP	P07491	777
FOLH1	MUC1	P13931	758
FOLH1	DPP4	P27487	758
FOLH1	MSLN	Q13421	755
FOLH1	TNFRSF9	Q07011	754

IntAct

9 interactions, top by confidence:

A	B	Type	Score
FOLH1	Dlg4	psi-mi:“MI:0407”(direct interaction)	0.440
FOLH1	MTAP	psi-mi:“MI:0915”(physical association)	0.370
RPS3	FOLH1	psi-mi:“MI:0915”(physical association)	0.370
MME		psi-mi:“MI:0914”(association)	0.350
TEX101	GGT3P	psi-mi:“MI:0914”(association)	0.350
SLC2A8	FOLH1	psi-mi:“MI:0914”(association)	0.350
FOLH1	GLG1	psi-mi:“MI:0914”(association)	0.350
LCOR	FOLH1	psi-mi:“MI:0915”(physical association)	0.000

BioGRID (18): CTH (Co-fractionation), FOLH1 (Affinity Capture-RNA), FOLH1 (Affinity Capture-Western), HDAC1 (Affinity Capture-Western), FOLH1 (Affinity Capture-Western), C3orf52 (Two-hybrid), DDX21 (Proximity Label-MS), FOLH1 (Affinity Capture-MS), FOLH1 (Affinity Capture-MS), GLG1 (Affinity Capture-MS), NAALAD2 (Affinity Capture-MS), FOLH1 (Affinity Capture-MS), FOLH1 (Two-hybrid), FOLH1 (Affinity Capture-MS), ITGB4 (Affinity Capture-Western)

ESM2 similar proteins: A3KG59, A4IFH5, B9N1F9, D3ZVR9, O04059, O35331, O35621, O46560, P11172, P24298, P37111, Q03154, Q04609, Q15124, Q17QK3, Q2R483, Q501L1, Q5E9T8, Q5I0K3, Q5NAY4, Q5R514, Q5R5C9, Q5RDE7, Q5RDN7, Q5RFB0, Q5RFI8, Q6AY30, Q6AYS7, Q6K2E8, Q6PTT0, Q6Q0N1, Q6ZV70, Q7TSV4, Q7X7L3, Q8BZF8, Q8CG45, Q8CG76, Q8IYS1, Q8K183, Q8N0X4

Diamond homologs: A0A1D6L709, B2GUY2, D4B1R0, O35409, O43023, O54697, O77564, P70627, P91406, Q04609, Q5RDH6, Q5WN23, Q7M758, Q7Y228, Q852M4, Q9CZR2, Q9HBA9, Q9JKX3, Q9M1S8, Q9UP52, Q9UQQ1, Q9Y3Q0, A4R017, A6REE0, B2W3C7, B2W572, B6K327, C4JHZ6, C5FP82, C9SPU8, D4AM42, D4D8N9, E3RJ99, E3S5D4, E4URG0, E5A6Z0, E5R501, Q01693, Q0UNS4, Q2U1F3

SIGNOR signaling

0 interactions.

Disease & clinical

Clinical variants and AI predictions

ClinVar

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

Classification	Count (floor)
Pathogenic	0
Likely pathogenic	0
Uncertain significance	64
Likely benign	10
Benign	4

Top pathogenic / likely-pathogenic (0)

SpliceAI

3311 predictions. Top by Δscore:

Variant	Effect	Δscore
11:49153844:A:AC	donor_gain	1.0000
11:49153845:C:CC	donor_gain	1.0000
11:49153845:CTTG:C	donor_gain	1.0000
11:49156714:TA:T	donor_loss	1.0000
11:49156715:A:AC	donor_gain	1.0000
11:49156715:A:T	donor_loss	1.0000
11:49156715:AC:A	donor_gain	1.0000
11:49156715:ACC:A	donor_gain	1.0000
11:49156716:C:CT	donor_gain	1.0000
11:49156716:CC:C	donor_gain	1.0000
11:49156716:CCC:C	donor_gain	1.0000
11:49156716:CCCA:C	donor_gain	1.0000
11:49156716:CCCAA:C	donor_gain	1.0000
11:49156803:TTATC:T	acceptor_gain	1.0000
11:49156804:TATC:T	acceptor_gain	1.0000
11:49156804:TATCC:T	acceptor_gain	1.0000
11:49156805:ATCCT:A	acceptor_gain	1.0000
11:49156806:TC:T	acceptor_gain	1.0000
11:49156806:TCC:T	acceptor_loss	1.0000
11:49156806:TCCTT:T	acceptor_gain	1.0000
11:49156807:CCTT:C	acceptor_gain	1.0000
11:49156808:C:CC	acceptor_gain	1.0000
11:49156808:C:T	acceptor_gain	1.0000
11:49156809:T:A	acceptor_loss	1.0000
11:49156809:T:C	acceptor_gain	1.0000
11:49156809:T:TC	acceptor_gain	1.0000
11:49156810:T:TC	acceptor_gain	1.0000
11:49156811:T:C	acceptor_gain	1.0000
11:49156811:T:TC	acceptor_gain	1.0000
11:49156817:A:AC	acceptor_gain	1.0000

AlphaMissense

4942 scored. Top likely-pathogenic:

Variant	Protein change	am_pathogenicity
11:49171242:A:G	W421R	0.999
11:49171242:A:T	W421R	0.999
11:49171243:G:C	S420R	0.998
11:49171243:G:T	S420R	0.998
11:49171245:T:G	S420R	0.998
11:49171236:C:G	A423P	0.997
11:49171240:C:A	W421C	0.996
11:49171240:C:G	W421C	0.996
11:49200368:A:G	W100R	0.996
11:49200368:A:T	W100R	0.996
11:49171261:T:A	R414S	0.995
11:49171261:T:G	R414S	0.995
11:49173441:A:G	W381R	0.995
11:49173441:A:T	W381R	0.995
11:49173447:C:G	D379H	0.995
11:49173449:C:G	R378P	0.995
11:49173467:A:T	V372D	0.995
11:49186656:G:C	F209L	0.995
11:49186656:G:T	F209L	0.995
11:49186658:A:G	F209L	0.995
11:49169214:A:C	N451K	0.994
11:49169214:A:T	N451K	0.994
11:49171215:C:G	G430R	0.994
11:49173446:T:A	D379V	0.993
11:49156781:T:A	D520V	0.992
11:49156782:C:G	D520H	0.992
11:49171211:G:A	S431F	0.992
11:49171214:C:T	G430D	0.992
11:49171262:C:G	R414T	0.992
11:49173412:A:C	S390R	0.992

dbSNP variants (sampled 300 via entrez): RS1000202032 (11:49170981 T>C), RS1000360408 (11:49177490 A>T), RS1000467076 (11:49183593 A>G), RS1000516092 (11:49191076 A>G), RS1000636387 (11:49146199 C>T), RS1000747212 (11:49197368 C>T), RS1000785326 (11:49153099 A>G), RS1000817978 (11:49184025 A>T), RS1000874288 (11:49204746 C>G), RS1000896133 (11:49160397 T>C), RS1000954461 (11:49166541 T>A), RS1001009793 (11:49166162 T>A,C), RS1001179831 (11:49196900 A>C), RS1001348529 (11:49160076 C>T), RS1001362467 (11:49179221 G>A)

Disease associations

OMIM: gene MIM:600934 | disease phenotypes:

GenCC curated gene-disease

Mondo (0):

Orphanet (0):

HPO phenotypes

0 total (0 of 0 shown, HPO-id order):

GWAS associations

22 associations (top):

Study	Trait	p-value
GCST000288_7	HDL cholesterol	6.000000e-11
GCST000817_142	Height	2.000000e-08
GCST005580_61	Intraocular pressure	3.000000e-12
GCST006585_880	Blood protein levels	3.000000e-26
GCST006947_4	Feeling fed-up	4.000000e-08
GCST007267_71	Systolic blood pressure	1.000000e-10
GCST008075_185	HDL cholesterol levels x alcohol consumption (regular vs non-regular drinkers) interaction (2df)	4.000000e-09
GCST008075_48	HDL cholesterol levels x alcohol consumption (regular vs non-regular drinkers) interaction (2df)	7.000000e-14
GCST008084_113	HDL cholesterol levels x alcohol consumption (drinkers vs non-drinkers) interaction (2df)	1.000000e-49
GCST008084_155	HDL cholesterol levels x alcohol consumption (drinkers vs non-drinkers) interaction (2df)	1.000000e-39
GCST008085_160	HDL cholesterol levels in current drinkers	5.000000e-09
GCST008085_185	HDL cholesterol levels in current drinkers	1.000000e-06
GCST009733_165	Urinary metabolite levels in chronic kidney disease	3.000000e-96
GCST010241_218	Apolipoprotein A1 levels	1.000000e-09
GCST010242_352	HDL cholesterol levels	8.000000e-11
GCST010250_1	Systolic blood pressure x dichotomous lifestyle risk score interaction (2df test)	7.000000e-07
GCST010989_99	Body size at age 10	3.000000e-08
GCST012020_178	Serum metabolite levels	3.000000e-84
GCST012020_179	Serum metabolite levels	4.000000e-58
GCST012021_103	Serum metabolite levels	3.000000e-84
GCST012021_104	Serum metabolite levels	4.000000e-58
GCST012353_37	Serum metabolite concentrations in chronic kidney disease	5.000000e-14

EFO canonical traits (9, from GWAS)

EFO ID	Trait name
EFO:0004612	high density lipoprotein cholesterol measurement
EFO:0004695	intraocular pressure measurement
EFO:0009588	feeling “fed-up” measurement
EFO:0006335	systolic blood pressure
EFO:0004329	alcohol drinking
EFO:0005116	urinary metabolite measurement
EFO:0004614	apolipoprotein A 1 measurement
EFO:0010724	lifestyle measurement
EFO:0009819	comparative body size at age 10, self-reported

Drugs & pharmacology

Drug and pharmacology data

Is drug target: yes

ChEMBL targets (1): CHEMBL1892 (SINGLE PROTEIN)

Molecules with ChEMBL bioactivity

7 molecules (phase ≥1), by development phase (incl. off-target/promiscuous compounds). Patent mentions across the top 20 by phase: 316,334 (via chembl_molecule»patent_compound — counts attach to the compound, not the gene–compound relationship, so off-target/promiscuous molecules can dominate).

Molecule	Name	Phase	Patents
CHEMBL3578202	GOZETOTIDE	4	466
CHEMBL4299851	PIFLUFOLASTAT	4	275
CHEMBL53463	DOXORUBICIN	4	314,282
CHEMBL3427435	CTT-1057	3	32
CHEMBL4594280	VIPIVOTIDE TETRAXETAN	2	971
CHEMBL496748	IOFOLASTAT	2	58
CHEMBL456029	MIP-1095	1	250

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

PharmGKB clinical annotations

1 annotations.

Variant	Type	Level	Drugs	Phenotypes
rs61886492	Toxicity	3	mercaptopurine;methotrexate	Acute lymphoblastic leukemia

PharmGKB variants

1 variants.

Variant	Genes	Level	Score	#Clin annots	Drugs
rs61886492	FOLH1	3	1.25	1	mercaptopurine;methotrexate

GtoPdb / IUPHAR curated pharmacology

(IUPHAR/BPS Guide to Pharmacology — expert-curated)

Target class: enzyme — M28: Aminopeptidase Y

Most potent curated ligand interactions (2 total), top 2:

Ligand	Action	Affinity	Parameter
vipivotide tetraxetan	Inhibition	9.43	pKi
compound 8d [PMID: 15027864]	Inhibition	8.52	pKi

Binding affinities (BindingDB)

167 measured of 186 human assays (187 total across all organisms); most potent 50 below. Values come from heterogeneous assays and are not directly comparable.

Ligand	Measure	Value	Patent
pentanedioic acid analogue, (S)-1	IC50	0.1 nM
US20250213705, Compound 120	KD	0.141 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 113	KD	0.142 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 102	KD	0.157 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 119	KD	0.161 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 109	KD	0.164 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 103	KD	0.169 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 2	KD	0.208 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 101	KD	0.249 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 110	KD	0.252 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 106	KD	0.288 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 108	KD	0.294 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 114	KD	0.294 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
pentanedioic acid analogue, (RS)-1	IC50	0.3 nM
CHEMBL2079731	KD	0.3 nM
US20250213705, Compound 104	KD	0.346 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 107	KD	0.379 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 112	KD	0.417 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 78	KD	0.43 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 68	KD	0.46 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 100	KD	0.477 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
CHEMBL2079727	KD	0.5 nM
US20250213705, Compound 87	KD	0.5 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 111	KD	0.512 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 115	KD	0.536 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 105	IC50	0.551 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 71	KD	0.56 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 12	KD	0.568 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 67	KD	0.6 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
(2S)-2-[[(1S)-1-carboxy-5-[6-[[1-(2-fluoroethyl)triazol-4-yl]methoxy]-2-oxo-3H-benzimidazol-1-yl]pentyl]carbamoylamino]pentanedioic acid	KI	0.63 nM	US-10894807: PSMA imaging agents
US20250213705, Compound 8	KD	0.654 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
Boc-AdamGPI Trimer	KD	0.7 nM
US20250213705, Compound 99	KD	0.73 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
5-[((S)-3-{3-[5-tert-Butoxycarbonylamino-3,7-bis-(2-{(S)-1-carboxy-3-[(2,4-dicarboxy-butyl)-hydroxy-phosphinoyl]-propylcarbamoyl}-ethyl)-adamantan-1-yl]-propionylamino}-3-carboxy-propyl)-hydroxy-phosphinoyl]-4-carboxy-pentanoic acid	KD	0.8 nM
US20250213705, Compound 34	KD	0.85 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 64	KD	0.92 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 74	KD	0.92 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 59	KD	0.98 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 69	KD	0.98 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 36	KD	0.996 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 92	KD	1.01 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 84	KD	1.05 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 37	KD	1.07 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 39	KD	1.08 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 61	KD	1.1 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 62	KD	1.11 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 52	KD	1.12 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 55	KD	1.15 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 56	KD	1.15 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE
US20250213705, Compound 86	KD	1.17 nM	US-20250213705: PSMA TARGETING LIGANDS AND METHODS OF USE

ChEMBL bioactivities

671 potent at pChembl≥5 of 693 total, top 50 by pChembl (potency: 10 = 0.1 nM, 6 = 1 µM).

pChembl	Type	Value	Unit	Molecule
11.00	Ki	0.01	nM	CHEMBL510924
11.00	Ki	0.01	nM	CHEMBL6007312
10.77	Ki	0.017	nM	CHEMBL5866602
10.74	Ki	0.018	nM	GOZETOTIDE
10.70	Ki	0.02	nM	CHEMBL5786057
10.68	IC50	0.021	nM	CHEMBL510924
10.52	Ki	0.03	nM	CHEMBL5994140
10.46	Ki	0.035	nM	CHEMBL5848906
10.44	Ki	0.036	nM	CHEMBL6010838
10.35	Ki	0.0449	nM	CHEMBL3309678
10.35	Ki	0.045	nM	CHEMBL3309678
10.31	Ki	0.049	nM	CHEMBL3578199
10.30	Ki	0.05	nM	CHEMBL510924
10.15	Ki	0.071	nM	CHEMBL3578197
10.15	Ki	0.07	nM	CHEMBL5786590
10.12	IC50	0.075	nM	CHEMBL4284995
10.04	IC50	0.091	nM	TUBULYSIN B
10.00	Ki	0.1	nM	CHEMBL189217
10.00	Ki	0.1	nM	CHEMBL5753520
9.96	Ki	0.11	nM	CHEMBL3578200
9.92	Ki	0.12	nM	CHEMBL5743096
9.89	IC50	0.13	nM	CHEMBL4280968
9.85	Ki	0.14	nM	CHEMBL4211875
9.82	IC50	0.15	nM	CHEMBL4286872
9.80	Ki	0.16	nM	CHEMBL5873627
9.77	Ki	0.17	nM	CHEMBL3311268
9.76	Ki	0.174	nM	CHEMBL3311268
9.71	Ki	0.194	nM	CHEMBL454897
9.70	Ki	0.2	nM	CHEMBL47009
9.70	IC50	0.2	nM	CHEMBL4288368
9.70	Kd	0.2	nM	CHEMBL2079731
9.70	Kd	0.2	nM	CHEMBL508430
9.70	Ki	0.2	nM	CHEMBL6015887
9.70	Ki	0.2	nM	CHEMBL5795495
9.70	Ki	0.2	nM	CHEMBL5798260
9.70	Ki	0.2	nM	CHEMBL6050284
9.66	Ki	0.221	nM	CHEMBL3309680
9.66	IC50	0.22	nM	CHEMBL510924
9.59	Ki	0.256	nM	CHEMBL454897
9.59	Ki	0.26	nM	CHEMBL5872050
9.58	Ki	0.265	nM	CHEMBL5876702
9.58	Ki	0.265	nM	CHEMBL5932456
9.57	Ki	0.268	nM	CHEMBL3309676
9.55	Ki	0.28	nM	CHEMBL47009
9.55	IC50	0.28	nM	CHEMBL4277729
9.52	IC50	0.3	nM	CHEMBL47009
9.52	Ki	0.3	nM	CHEMBL47009
9.52	Kd	0.3	nM	CHEMBL2079731
9.52	Kd	0.3	nM	CHEMBL508430
9.52	Ki	0.3	nM	CHEMBL189217

PubChem BioAssay actives

513 with measured affinity, of 821 total; 50 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
(2S)-2-[[(1S)-5-[[6-[4-[4-(4-acetylpiperazin-1-yl)phenyl]piperazin-1-yl]pyridine-3-carbonyl]-[(4-bromophenyl)methyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	<0.0001	uM
(2S)-2-[[(1S)-5-[(4-bromophenyl)methyl-[6-[4-[4-[4-[4-carboxy-3-(3-hydroxy-6-oxoxanthen-9-yl)benzoyl]piperazin-1-yl]phenyl]piperazin-1-yl]pyridine-3-carbonyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	<0.0001	uM
(2S)-2-[[(1S)-1-carboxy-5-[(4-iodobenzoyl)amino]pentyl]carbamoylamino]pentanedioic acid	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	<0.0001	uM
Gozetotide	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	<0.0001	uM
(2S)-2-[[(1S)-5-[acetyl-[(4-bromophenyl)methyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	<0.0001	uM
(2S)-2-(phosphonomethyl)pentanedioic acid	1797656: Microplate GCPII Radioactivity-Based Assay from Article 10.1021/jm049258g: “Enantiospecificity of glutamate carboxypeptidase II inhibition.”	ic50	0.0001	uM
(2S)-2-[[(1S)-5-[[6-[4-[4-(4-acetylpiperazin-1-yl)phenyl]piperazin-1-yl]pyridine-3-carbonyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	0.0001	uM
(2S)-2-[[(1S)-5-[[6-[4-[4-[4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyl]piperazin-1-yl]phenyl]piperazin-1-yl]pyridine-3-carbonyl]-[(4-bromophenyl)methyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1226540: Inhibition of antiTEV-tagged GCP2 extracellular portion (aa 44-750) (unknown origin) using folyl-di-L-glutamate as substrate by HPLC-based enzymatic assay	ki	0.0001	uM
(2S)-2-[[(1S)-1-carboxy-5-[(6-fluoro-3-pyridinyl)-methylamino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0001	uM
(2S)-2-[[(1S)-1-carboxy-5-[N-[2-[(6-fluoropyridine-3-carbonyl)amino]ethyl]anilino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0001	uM
3-[6-[[4-[[(2S)-1,5-bis[[(5S)-5-carboxy-5-[[(1S)-1,3-dicarboxypropyl]carbamoylamino]pentyl]amino]-1,5-dioxopentan-2-yl]amino]-4-oxobutyl]amino]-6-oxohexyl]-2-[(1E,3E,5E)-5-(3-ethyl-1,1-dimethyl-6,8-disulfobenzo[e]indol-2-ylidene)penta-1,3-dienyl]-1,1-dimethyl-8-sulfobenzo[e]indol-3-ium-6-sulfonate	1374082: Inhibition of PSMA in human LNCaP cell lysates incubated for 2 hrs in presence of N-acetylaspartylglutamate by Amplex red glutamic acid/glutamate oxidase assay	ki	0.0001	uM
(2S)-2-[[(1S)-1-carboxy-5-[N-[2-[(4-fluorobenzoyl)amino]ethyl]anilino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0001	uM
2-(phosphonomethyl)pentanedioic acid	74532: In vitro inhibitory activity against glutamate carboxypeptidase II (GCP II) using N-acetyl-L-aspartyl-[3H]L-glutamate as a substrate	ki	0.0002	uM
(2S)-2-[[(1S)-1-carboxy-5-(4-fluoro-N-methylanilino)-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0002	uM
(2S)-2-[[(1S)-1-carboxy-5-[(4-fluorobenzoyl)amino]pentyl]carbamoylamino]pentanedioic acid	409346: Inhibition of PSMA in human LNCaP cell extract using [3H]NAAG by fluorescence-based assay	ki	0.0002	uM
2-[[[(3S)-3-[3-[3,5-bis[3-[[(1S)-1-carboxy-3-[2,4-dicarboxybutyl(hydroxy)phosphoryl]propyl]amino]-3-oxopropyl]-7-[(2-methylpropan-2-yl)oxycarbonylamino]-1-adamantyl]propanoylamino]-3-carboxypropyl]-hydroxyphosphoryl]methyl]pentanedioic acid	411794: Binding affinity to PSMA in human PC3 cells assessed as cell surface binding in Tris-buffered saline buffer	kd	0.0002	uM
sodium 4-[[[(3S)-3-[3-[3,5-bis[3-[[(1S)-1-carboxy-3-[2,4-dicarboxybutyl(hydroxy)phosphoryl]propyl]amino]-3-oxopropyl]-7-[6-[2-[(E)-2-[(3E)-3-[(2E)-2-(3,3-dimethyl-1H-indol-2-ylidene)ethylidene]-2-(4-sulfonatophenoxy)cyclohexen-1-yl]ethenyl]-3,3-dimethylindol-1-ium-1-yl]hexanoylamino]-1-adamantyl]propanoylamino]-3-carboxypropyl]-hydroxyphosphoryl]methyl]-5-hydroxy-5-oxopentanoate	411795: Binding affinity to PSMA in human LNCaP cells assessed as cell surface binding in Tris-buffered saline buffer	kd	0.0002	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)-4-bromophenyl]carbamoylamino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0002	uM
(2S)-2-[[(1S)-5-acetamido-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0002	uM
(2S)-2-[[(1S)-5-[acetyl(benzyl)amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0003	uM
(2S)-2-[[(1S)-1-carboxy-5-(4-fluoroanilino)-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0003	uM
(2S)-2-[[(1S)-1-carboxy-5-(3-fluoroanilino)-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0003	uM
(2S)-2-[[[(2S)-2-carboxy-2-[[(4S)-4-carboxy-4-[6-[[3-carboxy-4-(3-hydroxy-6-oxoxanthen-9-yl)benzoyl]amino]hexanoylamino]butanoyl]amino]ethoxy]-hydroxyphosphoryl]amino]pentanedioic acid	631983: Inhibition of PSMA using N-[4-(phenylazo)-benzoyl]-glutamyl-gamma-glutamic acid as substrate after 15 mins by HPLC analysis	ic50	0.0003	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)-4-bromobenzoyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0003	uM
3-[6-[3-[bis[2-[4-[[(5S)-5-carboxy-5-[[(1S)-1,3-dicarboxypropyl]carbamoylamino]pentyl]carbamoyl]anilino]-2-oxoethyl]amino]propylamino]-6-oxohexyl]-2-[(1E,3E,5E)-5-(3-ethyl-1,1-dimethyl-6,8-disulfobenzo[e]indol-2-ylidene)penta-1,3-dienyl]-1,1-dimethyl-8-sulfobenzo[e]indol-3-ium-6-sulfonate	1374082: Inhibition of PSMA in human LNCaP cell lysates incubated for 2 hrs in presence of N-acetylaspartylglutamate by Amplex red glutamic acid/glutamate oxidase assay	ki	0.0004	uM
(2S)-6-[[3-[[(5S)-5-carboxy-5-[[(1S)-1-carboxy-2-[(2-hydroxy-3,4-dioxocyclobuten-1-yl)amino]ethyl]carbamoylamino]pentyl]carbamoyl]-5-fluorobenzoyl]amino]-2-[[(1S)-1-carboxy-2-[(2-hydroxy-3,4-dioxocyclobuten-1-yl)amino]ethyl]carbamoylamino]hexanoic acid	1989251: Binding affinity to human recombinant PSMA (unknown origin) assessed as inhibition constant incubated for 60 mins in presence of NAAG	ki	0.0004	uM
(2S)-6-[[3-[[(5S)-5-carboxy-5-[[(1S)-1-carboxy-2-[(2-hydroxy-3,4-dioxocyclobuten-1-yl)amino]ethyl]carbamoylamino]pentyl]carbamoyl]-5-iodobenzoyl]amino]-2-[[(1S)-1-carboxy-2-[(2-hydroxy-3,4-dioxocyclobuten-1-yl)amino]ethyl]carbamoylamino]hexanoic acid	1989251: Binding affinity to human recombinant PSMA (unknown origin) assessed as inhibition constant incubated for 60 mins in presence of NAAG	ki	0.0004	uM
(2S)-2-[[(1S)-1-carboxy-5-[(5-iodopyridine-3-carbonyl)amino]pentyl]carbamoylamino]pentanedioic acid	409345: Inhibition of PSMA in human LNCaP cell extract using [3H]NAAG by radiometric assay	ki	0.0004	uM
2-[[[(3S)-3-[3-[3,5-bis[3-[[(1S)-1-carboxy-3-[2,4-dicarboxybutyl(hydroxy)phosphoryl]propyl]amino]-3-oxopropyl]-7-[6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoylamino]-1-adamantyl]propanoylamino]-3-carboxypropyl]-hydroxyphosphoryl]methyl]pentanedioic acid	411795: Binding affinity to PSMA in human LNCaP cells assessed as cell surface binding in Tris-buffered saline buffer	kd	0.0004	uM
potassium 5-[[(1S)-1-carboxy-4-[[(1S)-1-carboxy-2-[[[(1S)-1,3-dicarboxypropyl]amino]-hydroxyphosphoryl]oxyethyl]amino]-4-oxobutyl]carbamoyl]-2-(3-hydroxy-6-oxoxanthen-9-yl)benzoate	631983: Inhibition of PSMA using N-[4-(phenylazo)-benzoyl]-glutamyl-gamma-glutamic acid as substrate after 15 mins by HPLC analysis	ic50	0.0004	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)phenyl]carbamoylamino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0004	uM
(2S)-2-[[(1S)-1-carboxy-3-methylbutyl]carbamoylamino]pentanedioic acid	1515290: Inhibition of N-terminally tagged human recombinant GCP2 (44 to 750 residues) extracellular domain expressed in Drosophila melanogaster S2 cells preincubated for 20 mins followed by [3H]-NAAG addition and measured after 20 mins by liquid scintillation counting assay	ic50	0.0005	uM
(2S)-2-[[(1S)-1-carboxy-5-[(3-fluorophenyl)methyl-methylamino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0005	uM
(2S)-2-[[(1S)-1-carboxy-5-[[4-[[[4-[[[4-[[[2-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetrazacyclododec-1-yl]acetyl]amino]methyl]benzoyl]amino]methyl]benzoyl]amino]methyl]benzoyl]amino]pentyl]carbamoylamino]pentanedioic acid	1949343: Binding affinity to PSMA in human LNCaP cells after 45 mins in presence of 68Ga-labeled PSMA-10 by gamma counting assay	ki	0.0005	uM
(2S)-2-[[(1S)-1-carboxy-5-[[(2S)-3-(4-phenylphenyl)-2-[[2-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetrazacyclododec-1-yl]acetyl]amino]propanoyl]amino]pentyl]carbamoylamino]pentanedioic acid	1949343: Binding affinity to PSMA in human LNCaP cells after 45 mins in presence of 68Ga-labeled PSMA-10 by gamma counting assay	ki	0.0005	uM
tetrasodium;4-[[[(3S)-3-[3-[3,5-bis[3-[[(1S)-1-carboxy-3-[2,4-dicarboxybutyl(hydroxy)phosphoryl]propyl]amino]-3-oxopropyl]-7-[6-[6-[2-[(E)-2-[(3E)-3-[(2E)-2-[3,3-dimethyl-5-sulfonato-1-(4-sulfonatobutyl)indol-2-ylidene]ethylidene]-2-(4-sulfonatophenoxy)cyclohexen-1-yl]ethenyl]-3,3-dimethyl-5-sulfonatoindol-1-ium-1-yl]hexanoylamino]hexanoylamino]-1-adamantyl]propanoylamino]-3-carboxypropyl]-hydroxyphosphoryl]methyl]-5-hydroxy-5-oxopentanoate	411795: Binding affinity to PSMA in human LNCaP cells assessed as cell surface binding in Tris-buffered saline buffer	kd	0.0005	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)benzoyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0006	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)-4-iodobenzoyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0006	uM
(2S)-2-[[(1S)-1-carboxy-3-methylbutyl]carbamoyloxy]pentanedioic acid	1515290: Inhibition of N-terminally tagged human recombinant GCP2 (44 to 750 residues) extracellular domain expressed in Drosophila melanogaster S2 cells preincubated for 20 mins followed by [3H]-NAAG addition and measured after 20 mins by liquid scintillation counting assay	ic50	0.0006	uM
3-[6-[[4-[[(5S)-5-carboxy-5-[[(1S)-1,3-dicarboxypropyl]carbamoylamino]pentyl]amino]-4-oxobutyl]amino]-6-oxohexyl]-2-[(1E,3E,5E)-5-(3-ethyl-1,1-dimethyl-6,8-disulfobenzo[e]indol-2-ylidene)penta-1,3-dienyl]-1,1-dimethyl-8-sulfobenzo[e]indol-3-ium-6-sulfonate	1374082: Inhibition of PSMA in human LNCaP cell lysates incubated for 2 hrs in presence of N-acetylaspartylglutamate by Amplex red glutamic acid/glutamate oxidase assay	ki	0.0006	uM
(2S)-2-[[(1S)-1-carboxy-5-[(4-fluorophenyl)methyl-methylamino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0006	uM
potassium [(2S)-2-[[(4S)-4-amino-4-carboxybutanoyl]amino]-2-carboxyethyl] [(1S)-1,3-dicarboxypropyl] phosphate	538387: Inhibition of PSMA in human LNCaP cells	ic50	0.0006	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)-4-chlorobenzoyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0007	uM
(4E)-4-[[1-[6-[[(5S)-5-carboxy-5-[[(1S)-1,3-dicarboxypropyl]carbamoylamino]pentyl]amino]-6-oxohexyl]-3,3-dimethylindol-1-ium-2-yl]methylidene]-2-[(Z)-[1-[6-[[(5S)-5-carboxy-5-[[(1S)-1,3-dicarboxypropyl]carbamoylamino]pentyl]amino]-6-oxohexyl]-3,3-dimethylindol-2-ylidene]methyl]-3-oxocyclobuten-1-olate	1989250: Binding affinity to PSMA (unknown origin) assessed as inhibition constant	ki	0.0007	uM
tetralithium;(2S)-2-(phosphonatoamino)pentanedioate	309919: Inhibition of PSMA	ic50	0.0009	uM
(2S)-2-[[(1S)-1-carboxy-5-[2-[(4-fluorobenzoyl)amino]ethylamino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0009	uM
(2S)-2-[[(1S)-1-carboxy-2-[(2-hydroxy-3,4-dioxocyclobuten-1-yl)amino]ethyl]carbamoylamino]-6-[(4-fluorobenzoyl)amino]hexanoic acid	1989251: Binding affinity to human recombinant PSMA (unknown origin) assessed as inhibition constant incubated for 60 mins in presence of NAAG	ki	0.0009	uM
(2S)-2-[[(1S)-1-carboxy-4-[(4-fluorobenzoyl)amino]butyl]carbamoylamino]pentanedioic acid	1989251: Binding affinity to human recombinant PSMA (unknown origin) assessed as inhibition constant incubated for 60 mins in presence of NAAG	ki	0.0009	uM
(2S)-2-[[(1S)-5-[[2-(2-acetamidoethoxy)-4-fluorobenzoyl]amino]-1-carboxypentyl]carbamoylamino]pentanedioic acid	1180461: Inhibition of human recombinant N-terminal Avi-tagged glutamate carboxypeptidase 2 extracellular domain (44 to 750 amino acids) using pteroyl-di-L-glutamate substrate by HPLC method	ki	0.0010	uM
(2S)-2-[[(1S)-1-carboxy-5-[(6-fluoro-3-pyridinyl)amino]-5-oxopentyl]carbamoylamino]pentanedioic acid	1423726: Inhibition of recombinant human PSMA using [3H]NAAG as substrate preincubated for 15 mins followed by substrate addition measured after 20 mins by liquid scintillation method	ic50	0.0010	uM

CTD chemical–gene interactions

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

Chemical	Actions (top 5)	PubMed papers
Metribolone	affects expression, decreases expression, decreases reaction, affects cotreatment	5
Cyclosporine	decreases expression, increases expression	3
Arsenic	affects response to substance, affects reaction, affects abundance, affects metabolic processing	2
tetramethrin	decreases expression	1
phentin acetate	increases expression	1
arsenite	increases methylation	1
tris(1,3-dichloro-2-propyl)phosphate	affects cotreatment, affects expression	1
sodium arsenite	decreases expression	1
monomethylarsonic acid	affects abundance, affects metabolic processing, affects response to substance	1
perfluorooctanoic acid	increases expression	1
isospaglumic acid	affects binding, decreases reaction	1
CGP 52608	affects binding, increases reaction	1
acyline	increases expression	1
difenoconazole	increases expression	1
bombesin (7-14)	affects binding, decreases reaction	1
2,2’,4,4’-tetrabromodiphenyl ether	increases expression	1
1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane	decreases reaction, increases expression, decreases expression	1
2-(3-(1,3-dicarboxypropyl)ureido)pentanedioic acid	decreases reaction, affects binding	1
1-(1,3-carboxypropyl)-4,7-carboxymethyl-1,4,7-triazacyclononane	decreases reaction, affects binding	1
Benzo(a)pyrene	decreases methylation, increases methylation	1
Calcitriol	affects reaction, decreases expression	1
Copper	affects binding, decreases reaction	1
Dichlorodiphenyl Dichloroethylene	decreases reaction, increases expression, decreases expression	1
DDT	decreases expression, decreases reaction, increases expression	1
Endosulfan	affects cotreatment, decreases expression	1
Nickel	decreases expression	1
Oxygen	increases expression	1
Pentobarbital	increases response to substance	1
Selenium	affects cotreatment, decreases expression	1
Silicon Dioxide	decreases expression	1

ChEMBL screening assays

139 unique, capped per target: 131 binding, 7 admet, 1 functional

Representative assays (with source publication via chembl_document):

Assay ID	Type	Description	Source paper
CHEMBL1020694	Binding	Binding affinity to PSMA in human PC3 cells assessed as cell surface binding in Tris-buffered saline buffer	Multivalent scaffolds for affinity maturation of small molecule cell surface binders and their application to prostate tumor targeting. — J Med Chem
CHEMBL4393043	ADMET	Binding affinity to PSMA in human LNCAP cells assessed as doxorubicin release in cytoplasm measured after 48 hrs	Synthesis and biological evaluation of Doxorubicin-containing conjugate targeting PSMA. — Bioorg Med Chem Lett
CHEMBL765765	Functional	Concentration required for inhibition of 50% of the cleavage of para-nitroanilide from MeO-Suc-Arg-Pro-Tyr-pNA.HCl by Prostate specific antigen PSA	Design and synthesis of novel monocyclic -lactam inhibitors of prostate specific antigen — Bioorg Med Chem Lett

Cellosaurus cell lines

8 cell lines: 6 cancer cell line, 1 spontaneously immortalized cell line, 1 transformed cell line

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

Cellosaurus	Name	Category	Sex
CVCL_B8G7	Abcam HCT 116 FOLH1 KO	Cancer cell line	Male
CVCL_B9IG	Abcam A-549 FOLH1 KO	Cancer cell line	Male
CVCL_D3N6	PC-3 9-7-11	Cancer cell line	Male
CVCL_E6QG	Genomeditech CHO-K1 H_FOLH1(PSMA)	Spontaneously immortalized cell line	Female
CVCL_E9XK	PC3-PIP	Cancer cell line	Male
CVCL_E9XY	PC3-FOLH1	Cancer cell line	Male
CVCL_SN83	HAP1 FOLH1 (-)	Cancer cell line	Male
CVCL_UE40	293T human FOLH1	Transformed cell line	Female

Clinical trials (associated diseases)

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