CARD8

Summary

CARD8 (caspase recruitment domain family member 8, HGNC:17057) is a protein-coding gene on chromosome 19q13.33, encoding Caspase recruitment domain-containing protein 8 (Q9Y2G2). Inflammasome sensor, which mediates inflammasome activation in response to various pathogen-associated signals, leading to subsequent pyroptosis of CD4(+) T-cells and macrophages.

The protein encoded by this gene belongs to the caspase recruitment domain (CARD)-containing family of proteins, which are involved in pathways leading to activation of caspases or nuclear factor kappa-B (NFKB). This protein may be a component of the inflammasome, a protein complex that plays a role in the activation of proinflammatory caspases. It is thought that this protein acts as an adaptor molecule that negatively regulates NFKB activation, CASP1-dependent IL1B secretion, and apoptosis. Polymorphisms in this gene may be associated with a susceptibility to rheumatoid arthritis. Alternatively spliced transcript variants have been described for this gene.

Source: NCBI Gene 22900 — RefSeq curated summary.

At a glance

• Gene–disease (curated): inflammatory bowel disease 30 (Limited, GenCC)
• Clinical variants (ClinVar): 498 total — 1 likely-pathogenic
• Phenotypes (HPO): 13
• MANE Select transcript: NM_001184900

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:17057
Approved symbol	CARD8
Name	caspase recruitment domain family member 8
Location	19q13.33
Locus type	gene with protein product
Status	Approved
Aliases	TUCAN, KIAA0955, CARDINAL, NDPP, Dakar
Ensembl gene	ENSG00000105483
Ensembl biotype	protein_coding
OMIM	609051
Entrez	22900

Gene structure

Transcript identifiers

Ensembl transcripts: 49 — 31 protein_coding, 9 nonsense_mediated_decay, 5 retained_intron, 4 protein_coding_CDS_not_defined

ENST00000377461, ENST00000391898, ENST00000447740, ENST00000517510, ENST00000517778, ENST00000518450, ENST00000518596, ENST00000518622, ENST00000518979, ENST00000519302, ENST00000519332, ENST00000519646, ENST00000519940, ENST00000520007, ENST00000520015, ENST00000520153, ENST00000520753, ENST00000521092, ENST00000521415, ENST00000521437, ENST00000521613, ENST00000522051, ENST00000522068, ENST00000522431, ENST00000522773, ENST00000522889, ENST00000523579, ENST00000523668, ENST00000523750, ENST00000600800, ENST00000651546, ENST00000877775, ENST00000877776, ENST00000877777, ENST00000877778, ENST00000877779, ENST00000877780, ENST00000877781, ENST00000877782, ENST00000877783, ENST00000877784, ENST00000877785, ENST00000930774, ENST00000930775, ENST00000946857, ENST00000946858, ENST00000946859, ENST00000946860, ENST00000946861

RefSeq mRNA: 18 — MANE Select: NM_001184900 NM_001184900, NM_001184901, NM_001184902, NM_001184903, NM_001184904, NM_001351782, NM_001351783, NM_001351784, NM_001351786, NM_001351787, NM_001351788, NM_001351789, NM_001351790, NM_001351791, NM_001351792, NM_001365950, NM_001426741, NM_014959

CCDS: CCDS12712, CCDS54287, CCDS54288, CCDS54289

Canonical transcript exons

ENST00000651546 — 14 exons

Exon	Start	End
ENSE00001754537	48249523	48249633
ENSE00002121739	48249751	48249847
ENSE00003465314	48238383	48238532
ENSE00003470054	48230777	48231006
ENSE00003495673	48230438	48230700
ENSE00003566731	48240962	48241063
ENSE00003574024	48232453	48232493
ENSE00003575909	48218871	48219012
ENSE00003606074	48221730	48221855
ENSE00003639465	48215340	48215384
ENSE00003640346	48231660	48231810
ENSE00003651170	48234403	48234543
ENSE00003841269	48208086	48211975
ENSE00003850926	48255792	48255912

Expression profiles

Bgee: expression breadth ubiquitous, 260 present calls, max score 97.04.

FANTOM5 (CAGE): breadth ubiquitous, TPM avg 17.4390 / max 443.4716, expressed in 1735 samples.

FANTOM5 promoters (6 alternative TSS)

Promoter ID	TPM avg	Samples expressed
181823	7.5664	615
181827	5.3731	1669
181825	2.0648	409
181824	1.5728	428
181826	0.7293	369
181822	0.1326	72

Top tissues by expression

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

Tissue	Anatomy ID	Expression score	Quality
monocyte	CL:0000576	97.04	gold quality
mononuclear cell	CL:0000842	96.87	gold quality
leukocyte	CL:0000738	96.76	gold quality
granulocyte	CL:0000094	96.16	gold quality
spleen	UBERON:0002106	95.87	gold quality
buccal mucosa cell	CL:0002336	95.82	gold quality
blood	UBERON:0000178	95.59	gold quality
lymph node	UBERON:0000029	94.18	gold quality
omental fat pad	UBERON:0010414	93.70	gold quality
gall bladder	UBERON:0002110	93.69	gold quality
peritoneum	UBERON:0002358	93.63	gold quality
adipose tissue of abdominal region	UBERON:0007808	93.43	gold quality
vermiform appendix	UBERON:0001154	93.40	gold quality
calcaneal tendon	UBERON:0003701	93.06	gold quality
right lung	UBERON:0002167	93.01	gold quality
bone marrow cell	CL:0002092	92.55	gold quality
colonic epithelium	UBERON:0000397	92.51	gold quality
ventricular zone	UBERON:0003053	92.49	gold quality
upper lobe of left lung	UBERON:0008952	92.39	gold quality
upper lobe of lung	UBERON:0008948	91.82	gold quality
small intestine Peyer’s patch	UBERON:0003454	91.72	gold quality
apex of heart	UBERON:0002098	91.44	gold quality
subcutaneous adipose tissue	UBERON:0002190	91.27	gold quality
adipose tissue	UBERON:0001013	91.07	gold quality
bone marrow	UBERON:0002371	90.91	gold quality
connective tissue	UBERON:0002384	90.66	gold quality
rectum	UBERON:0001052	90.57	gold quality
caecum	UBERON:0001153	90.49	gold quality
body of uterus	UBERON:0009853	90.41	gold quality
right lobe of thyroid gland	UBERON:0001119	90.23	gold quality

Single-cell (SCXA)

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

Experiment	Marker?	Max mean expression
E-ANND-3	yes	8.47

Regulation

Is transcription factor: no

Upstream regulators (CollecTRI, top): NFKB1, NFKB, RELA

miRNA regulators (miRDB)

141 targeting CARD8, 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-3613-3P	100.00	76.36	7965
HSA-MIR-5692A	100.00	74.40	6850
HSA-MIR-4262	100.00	73.26	3931
HSA-MIR-340-5P	100.00	72.50	4437
HSA-MIR-3662	99.99	73.82	5684
HSA-MIR-371B-5P	99.99	75.34	4759
HSA-MIR-4282	99.99	75.36	6408
HSA-MIR-548C-3P	99.99	74.01	7587
HSA-MIR-3692-3P	99.98	70.27	2139
HSA-MIR-4803	99.98	71.99	3117
HSA-MIR-373-5P	99.98	75.36	4753
HSA-MIR-616-5P	99.98	75.58	4775
HSA-MIR-548P	99.98	72.25	3784
HSA-MIR-12136	99.98	72.81	5713
HSA-MIR-548N	99.98	71.94	4170
HSA-MIR-3148	99.97	75.06	6478
HSA-MIR-3688-3P	99.97	72.02	2834
HSA-MIR-548AJ-3P	99.96	73.38	5345
HSA-MIR-548X-3P	99.96	73.38	5345
HSA-MIR-570-3P	99.96	72.41	4910
HSA-MIR-3605-5P	99.96	67.12	932
HSA-MIR-1250-3P	99.96	70.04	4038
HSA-MIR-23A-3P	99.95	74.24	3163
HSA-MIR-23B-3P	99.95	74.24	3163
HSA-MIR-23C	99.95	73.92	3192
HSA-MIR-545-3P	99.95	70.74	2783
HSA-MIR-548J-3P	99.94	72.61	4881
HSA-MIR-141-3P	99.94	72.79	2421
HSA-MIR-200A-3P	99.94	72.68	2420
HSA-MIR-548AE-3P	99.93	72.66	4867

Literature-anchored findings (GeneRIF, showing 40)

• CARD-8 protein, a new CARD family member that regulates caspase-1 activation and apoptosis. (card-8 protein) (PMID:11821383)
• Expression and characterization of NDPP1 (PMID:11956601)
• TUCAN/CARDINAL and DRAL participate in a common pathway for modulation of NF-kappaB activation. (PMID:12067710)
• TUCAN does not play a role in inhibition of procaspase-9 and in determining the sensitivity to cisplatin in Non Small Cells Lung Cancer. (PMID:16796750)
• association between a likely functional polymorphism in TUCAN and Crohn disease (PMID:17030188)
• no significant association between the risk allele “A” at Cys10Stop and risk for Crohn’s disease or ulcerative colitis was detected in patients of German and Norwegian descent (PMID:17484912)
• deleterious polymorphism of CARD8 may help predict the severity of rheumatoid arthritis (PMID:17878386)
• Analyzing 3 independent European IBD cohorts, we found no evidence that the C10X variant in CARD8 confers susceptibility for CD. (PMID:18092344)
• isoforms of CARD8 differ in their N-termini, resulting in diverse predicted molecular weights (47, 48, 51, 54 and 60 kDa) and multiple outcomes for the variant including Cys10Stop, Cys34Stop, Phe52Ile and Phe102Ile (PMID:18212821)
• The NALP3 and TUCAN single-nucleotide polymorphisms may explain the increased IL-1beta levels and inflammatory symptoms observed, but further studies are needed to reveal a functional relationship. (PMID:18311798)
• Women, but not men, carrying the CARD8 AA genotype (truncated protein) had a 2.39-fold higher risk of developing Alzheimer’s disease than subjects with the CARD8 TT genotype (full-length protein) (PMID:18841008)
• Gene-gene interaction between CARD8 and interleukin-6 reduces Alzheimer’s disease risk. (PMID:19252766)
• CARD8 and NALP3 genes combined polymorphism has a role of developing Crohn disease in men. (PMID:19319132)
• Carriage of CARD8-X is associated with a worse disease course in early rheumatoid arthritis. (PMID:19443463)
• Variation in the innate immunity genes CARD4, CARD8 and CARD15 is unlikely to play a major role in the susceptibility to CRC in the German population. (PMID:19843337)
• Study shows an essential role for apoptosis signal-regulating kinase 1 (ASK1), together with both c-jun-N-terminal kinase (JNK) and p38 pathways, and caspase-8 in Fas-induced apoptosis. (PMID:19940360)
• propose that CARD8-NALP3 genotype combinations protect against gut inflammation by preventing the NALP3 inflammasome from producing excessive interleukin-1be (PMID:20182451)
• Data show that CARD8 represents a novel molecular switch involved in the endogenous regulation of NOD2-dependent inflammatory processes in epithelial cells. (PMID:20385562)
• CARD8 variants might have roles in the pathogenesis of Crohn’s disease and ulcerative colitis in Koreans (PMID:21248762)
• Caspase8 polymorphism IVS12-19 G>A but not CASP8 -652 6N del polymorphism may modulate risk of esophageal squamous cell carcinoma and its survival outcome in northern Indian population. (PMID:21308686)
• analysis of interaction of the inflammasome genes CARD8 and NLRP3 in abdominal aortic aneurysms (PMID:21621776)
• results identify a function for the FIIND and show that CARD8 and NLRP1 are ZU5-UPA domain-containing autoproteolytic proteins, thus suggesting a novel mechanism for regulating innate immune responses (PMID:22087307)
• Variations in the CARD8 and NLRP3 genes are not associated with rheumatoid arthritis in the French as well general Tunisian population. (PMID:22128899)
• Twelve single nucleotide polymorphisms within NLRP1, NLRP3, NLRC4, CARD8, CASP1, and IL1B genes were analyzed in 150 HIV-1-infected Brazilian subjects. (PMID:22227487)
• we performed a genetic association study in patients with pneumococcal meningitis and found that single-nucleotide polymorphisms in the inflammasome genes CARD8 and NLRP1 are associated with poor disease outcome. (PMID:23053059)
• Results suggest that the CARD8 rs2043211 gene variant does not in susceptibility to rheumatoid arthritis (RA) or in the development of cardiovascular disease in patients with RA. (PMID:23088220)
• Mutation in the CARD8, a component of inflammasome, is associated with lower levels of antibodies directed to mannans and glucans at least in Crohn’s disease patients. (PMID:23506543)
• The results of this study support the novel association between CARD8 gene and HIV+tuberculosis coinfection, demonstrating that inflammasome genetics could influence HIV-1 infection and the development of opportunistic infection. (PMID:23507658)
• In a Swedish population, the minor allele of CARD8-C10X is associated with a decreased risk of AS, but not with levels of faecal calprotectin or disease phenotype. (PMID:23547871)
• Genetic variation in the inflammasome affects atopic dermatitis susceptibility. (PMID:23563199)
• Data indicate that CARD8 (caspase recruitment domain 8) mRNA was highly expressed in atherosclerotic plaques, and the minor allele was associated with lower expression of CARD8 in the plaques, suggesting that CARD8 may promote inflammation. (PMID:23611467)
• The first crystal structure of the CARD8 caspase-recruitment domain is reported. It adopts a six-helix bundle fold with a unique conformation of the alpha6 helix that is described for the first time. (PMID:23695559)
• levels of inflammasome-produced cytokines as a measure of inflammasome activation in healthy individuals carrying Q705K polymorphism in the NLRP3 gene combined with C10X in the CARD8 gene (PMID:24098386)
• ANRIL may increase the risk of ischemic stroke through regulation of the CARD8 pathway (PMID:24385277)
• we show that CARD8 plays a role as a negative regulator of NLRP3 inflammasome through its binding with NLRP3 (PMID:24517500)
• genetic polymorphism is associated with susceptibility to Crohn’s disease under the dominant model and homozygote contrast in the European population; meta-analysis (PMID:25564880)
• Patients carrying genotype TT of CARD8 rs2043211 polymorphism had higher triglycerides levels compared to those carrying the AA genotype. (PMID:25790751)
• The polymorphism of rs2043211 in CARD8 may be a relevant host susceptibility factor for the development of preeclampsia in the Chinese Han population. (PMID:25895569)
• CARD8 might not play a role in the pathogenesis of Tourette syndrome in Chinese Han population (PMID:25921775)
• Data indicate 3 variants in 3 novel genes myc target 1 protein (MYCT1), caspase recruitment domain family member 8 (CARD8) and zinc finger protein 543 (ZNF543), associated with familial IgA nephropathy (IgAN). (PMID:26095808)

Cross-species orthologs

0 orthologs

Protein

Protein identifiers

Caspase recruitment domain-containing protein 8 — Q9Y2G2 (reviewed: Q9Y2G2)

Alternative names: CARD-inhibitor of NF-kappa-B-activating ligand, Tumor up-regulated CARD-containing antagonist of CASP9

All UniProt accessions (10): Q9Y2G2, E5RFI5, E5RGC9, E5RGG3, E5RHJ3, E5RHZ3, E5RIN1, E5RJG3, H0YB90, H0YBW7

UniProt curated annotations — full annotation on UniProt →

Function. Inflammasome sensor, which mediates inflammasome activation in response to various pathogen-associated signals, leading to subsequent pyroptosis of CD4(+) T-cells and macrophages. Inflammasomes are supramolecular complexes that assemble in the cytosol in response to pathogens and other damage-associated signals and play critical roles in innate immunity and inflammation. Acts as a recognition receptor (PRR): recognizes specific pathogens and other damage-associated signals, such as HIV-1 protease activity or Val-boroPro inhibitor, and mediates CARD8 inflammasome activation. In response to pathogen-associated signals, the N-terminal part of CARD8 is degraded by the proteasome, releasing the cleaved C-terminal part of the protein (Caspase recruitment domain-containing protein 8, C-terminus), which polymerizes to initiate the formation of the inflammasome complex: the CARD8 inflammasome directly recruits pro-caspase-1 (proCASP1) independently of PYCARD/ASC and promotes caspase-1 (CASP1) activation, which subsequently cleaves and activates inflammatory cytokines IL1B and IL18 and gasdermin-D (GSDMD), leading to pyroptosis. Ability to sense HIV-1 protease activity leads to the clearance of latent HIV-1 in patient CD4(+) T-cells after viral reactivation; in contrast, HIV-1 can evade CARD8-sensing when its protease remains inactive in infected cells prior to viral budding. Also acts as a negative regulator of the NLRP3 inflammasome. May also act as an inhibitor of NF-kappa-B activation. Constitutes the precursor of the CARD8 inflammasome, which mediates autoproteolytic processing within the FIIND domain to generate the N-terminal and C-terminal parts, which are associated non-covalently in absence of pathogens and other damage-associated signals. Regulatory part that prevents formation of the CARD8 inflammasome: in absence of pathogens and other damage-associated signals, interacts with the C-terminal part of CARD8 (Caspase recruitment domain-containing protein 8, C-terminus), preventing activation of the CARD8 inflammasome. In response to pathogen-associated signals, this part is ubiquitinated by the N-end rule pathway and degraded by the proteasome, releasing the cleaved C-terminal part of the protein, which polymerizes and forms the CARD8 inflammasome. Constitutes the active part of the CARD8 inflammasome. In absence of pathogens and other damage-associated signals, interacts with the N-terminal part of CARD8 (Caspase recruitment domain-containing protein 8, N-terminus), preventing activation of the CARD8 inflammasome. In response to pathogen-associated signals, the N-terminal part of CARD8 is degraded by the proteasome, releasing this form, which polymerizes to form the CARD8 inflammasome complex: the CARD8 inflammasome complex then directly recruits pro-caspase-1 (proCASP1) and promotes caspase-1 (CASP1) activation, leading to gasdermin-D (GSDMD) cleavage and subsequent pyroptosis.

Subunit / interactions. Interacts with DPP9; leading to inhibit activation of the inflammasome. DPP9 acts via formation of a ternary complex, composed of a DPP9 homodimer, one full-length CARD8 protein, and one cleaved C-terminus of CARD8 (Caspase recruitment domain-containing protein 8, C-terminus). Interacts with DPP8; leading to inhibit activation of the inflammasome, probably via formation of a ternary complex with DPP8. Interacts with NLRP3. Interacts with IKBKG/NEMO. Interacts with DRAL. Binds to caspase-1 (CASP1), CARD16/pseudo-ICE and CARD18/ICEBERG. Interacts with NLRP2 (via NACHT domain). Interacts with the C-terminal part of CARD8 (Caspase recruitment domain-containing protein 8, C-terminus) in absence of pathogens and other damage-associated signals. Interacts with the N-terminal part of CARD8 (Caspase recruitment domain-containing protein 8, N-terminus) in absence of pathogens and other damage-associated signals. Homomultimer; forms the CARD8 inflammasome polymeric complex, a filament composed of homopolymers of this form in response to pathogens and other damage-associated signals. The CARD8 inflammasome polymeric complex directly recruits pro-caspase-1 (proCASP1) independently of PYCARD/ASC. Interacts (via CARD domain) with CASP1 (via CARD domain); leading to CASP1 activation.

Subcellular location. Cytoplasm. Nucleus Inflammasome.

Tissue specificity. High expression in lung, ovary, testis and placenta. Lower expression in heart, kidney and liver. Also expressed in spleen, lymph node and bone marrow.

Post-translational modifications. Undergoes autocatalytic processing within the FIIND domain to generate the N-terminal and C-terminal parts, which are associated non-covalently in absence of pathogens and other damage-associated signals. Ubiquitinated by the N-end rule pathway in response to pathogens and other damage-associated signals, leading to its degradation by the proteasome and subsequent release of the cleaved C-terminal part of the protein (Caspase recruitment domain-containing protein 8, C-terminus), which polymerizes and forms the CARD8 inflammasome. (Microbial infection) Proteolytic cleavage by HIV-1 protease in the disordered region and within the ZU5 region of the FIIND domain promotes ubiquitination of the N-terminal part by the N-end rule pathway and degradation by the proteasome, releasing the cleaved C-terminal part of the protein (Caspase recruitment domain-containing protein 8, C-terminus), which polymerizes and forms the CARD8 inflammasome. Undergoes less autocatalytic processing within the FIIND domain compared to isoform 5.

Disease relevance. Inflammatory bowel disease 30 (IBD30) [MIM:619079] A chronic, relapsing inflammation of the gastrointestinal tract with a complex etiology and a multifactorial inheritance pattern. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may affect any part of the gastrointestinal tract from the mouth to the anus, but most frequently it involves the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints. The disease may be caused by variants affecting the gene represented in this entry. A number of groups have studied the possible association between variant rs2043211 and inflammatory bowel disease. According to some studies involving a limited number of patients, this variant is associated with inflammatory bowel disease. Such association is however not confirmed in studies involving a large number of patients. Discrepancies between studies may be caused by the variable consequences of this polymorphism in the different isoforms. Whereas rs2043211 introduces a stop codon after ‘Cys-10’ (Cys10Ter) in isoform 1, and therefore the likely formation of a downstream transcriptional start site for this isoform, it causes Ile-102 variation in isoform 5, due to the upstream start site. Moreover, most patients bearing this polymorphism continue to express the slightly smaller but fully functional isoform 7, as a result of transcription downstream of the rs2043211 polymorphism.

Activity regulation. CARD8 inflammasome is activated by HIV-1 protease activity: HIV-1 protease cleaves CARD8, promoting ubiquitination and degradation of the N-terminal part, releasing the cleaved C-terminal part of the protein (Caspase recruitment domain-containing protein 8, C-terminus), which polymerizes and forms the CARD8 inflammasome. CARD8 inflammasome is inhibited by DPP8 and DPP9, which sequester the C-terminal fragment of CARD8 (Caspase recruitment domain-containing protein 8, C-terminus) in a ternary complex, thereby preventing CARD8 oligomerization and activation. CARD8 inflammasome is activated by Val-boroPro (Talabostat, PT-100), an inhibitor of dipeptidyl peptidases DPP8 and DPP9. Val-boroPro relieves inhibition of DPP8 and/or DPP9 by inducing the proteasome-mediated destruction of the N-terminal part of CARD8, releasing its C-terminal part from autoinhibition. Indirectly activated by the pseudodipeptide CQ31. CQ31 directly inactivates the peptidases PEPD and XPNPEP1, leading to an accumulation of dipeptides that weaky inhibit DDP8 and DPP9, relieving DPP8- and/or DPP9-mediated inhibition of CARD8.

Domain organisation. The disordered region is required for activation of the CARD8 inflammasome. The C-terminal part of CARD8 oligomerizes to form the core of the CARD8 inflammasome filament: in the filament, the CARD domains form a central helical filaments that are promoted by oligomerized, but flexibly linked, UPA regions surrounding the filaments. The UPA region reduces the threshold needed for filament formation and signaling. Directly recruits and polymerizes with the CARD domain of caspase-1 (CASP1) through the favorable side of the growing filament seed.

Isoforms (7)

UniProt ID	Names	Canonical?
Q9Y2G2-5	5, T60, a	yes
Q9Y2G2-1	1, Long, T48
Q9Y2G2-2	2, Short
Q9Y2G2-3	3, c
Q9Y2G2-4	4, T54, b
Q9Y2G2-6	6, d
Q9Y2G2-7	7, T47

RefSeq proteins (18): NP_001171829, NP_001171830, NP_001171831, NP_001171832, NP_001171833, NP_001338711, NP_001338712, NP_001338713, NP_001338715, NP_001338716, NP_001338717, NP_001338718, NP_001338719, NP_001338720, NP_001338721, NP_001352879, NP_001413670, NP_055774 (=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR001315	CARD	Domain
IPR011029	DEATH-like_dom_sf	Homologous_superfamily
IPR025307	FIIND_dom	Domain
IPR051249	NLRP_Inflammasome	Family

Pfam: PF00619, PF13553, PF23679

UniProt features (103 total): mutagenesis site 36, strand 20, helix 12, sequence conflict 8, splice variant 7, sequence variant 5, region of interest 4, chain 3, turn 3, site 2, domain 2, compositionally biased region 1

Structure

Experimental structures (PDB)

5 structures.

PDB	Method	Resolution (Å)
4IKM	X-RAY DIFFRACTION	2.46
7JKQ	ELECTRON MICROSCOPY	3.3
7JN7	ELECTRON MICROSCOPY	3.3
6XKJ	ELECTRON MICROSCOPY	3.54
6K9F	ELECTRON MICROSCOPY	3.7

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-Q9Y2G2-F1	71.42	0.42

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 (2): 59–60 ((microbial infection) cleavage; by hiv-1 protease); 296–297 (cleavage; by autolysis)

Mutagenesis-validated functional residues (36):

Position	Phenotype
152	increased autocalalytic cleavage.
51	does not affect cleavage by hiv-1 protease.
52	does not affect cleavage by hiv-1 protease.
53	does not affect cleavage by hiv-1 protease.
54	does not affect cleavage by hiv-1 protease.
55	does not affect cleavage by hiv-1 protease.
56	does not affect cleavage by hiv-1 protease.
57	does not affect cleavage by hiv-1 protease.
58	does not affect cleavage by hiv-1 protease.
59	abolished cleavage by hiv-1 protease, leading to prevent formation of the card8 inflammasome and subsequent pyroptosis.
60	abolished cleavage by hiv-1 protease, leading to prevent formation of the card8 inflammasome and subsequent pyroptosis.
157	does not affect sensitivity to val-boropro.
240	no effect on autocatalytic cleavage.
242	no effect on autocatalytic cleavage.
252	severe loss of autocatalytic cleavage.
270	severe loss of autocatalytic cleavage.
274	abolished interaction with dpp9, without affecting autocatalytic cleavage.
279	partial loss of autocatalytic cleavage.
280	no effect on autocatalytic cleavage.
295	partial loss of autocatalytic cleavage.
295	no effect on autocatalytic cleavage.
296	severe loss of autocatalytic cleavage.
297	complete loss of autocatalytic cleavage. abolished ability to form the card8 inflammasome and trigger pyroptosis. abolis
333	no effect on autocatalytic cleavage.
368	does not affect autocatalytic cleavage; does not affect interaction with dpp9; impaired interaction with the c-terminal

Function

Pathways and Gene Ontology

Reactome pathways

7 pathways

ID	Pathway
R-HSA-111458	Formation of apoptosome
R-HSA-9627069	Regulation of the apoptosome activity
R-HSA-109581	Apoptosis
R-HSA-109606	Intrinsic Pathway for Apoptosis
R-HSA-111461	Cytochrome c-mediated apoptotic response
R-HSA-111471	Apoptotic factor-mediated response
R-HSA-5357801	Programmed Cell Death

MSigDB gene sets: 241 (showing top): GOBP_NEGATIVE_REGULATION_OF_PROTEIN_CONTAINING_COMPLEX_ASSEMBLY, GOBP_NEGATIVE_REGULATION_OF_INTERLEUKIN_1_PRODUCTION, GOBP_INFLAMMATORY_RESPONSE, GOBP_RESPONSE_TO_PEPTIDE, GOBP_CELLULAR_RESPONSE_TO_LIPID, GOBP_CELLULAR_RESPONSE_TO_BIOTIC_STIMULUS, GOBP_CANONICAL_NF_KAPPAB_SIGNAL_TRANSDUCTION, GOBP_POSITIVE_REGULATION_OF_CYTOKINE_PRODUCTION, GOBP_NEGATIVE_REGULATION_OF_TUMOR_NECROSIS_FACTOR_MEDIATED_SIGNALING_PATHWAY, GOBP_REGULATION_OF_CELLULAR_COMPONENT_BIOGENESIS, GOBP_CELLULAR_RESPONSE_TO_OXYGEN_CONTAINING_COMPOUND, GOBP_INTERLEUKIN_1_PRODUCTION, GOBP_NEGATIVE_REGULATION_OF_INTRACELLULAR_SIGNAL_TRANSDUCTION, GOBP_NEGATIVE_REGULATION_OF_CELLULAR_COMPONENT_ORGANIZATION, GOBP_NEGATIVE_REGULATION_OF_RESPONSE_TO_BIOTIC_STIMULUS

GO Biological Process (22): activation of innate immune response (GO:0002218), inflammatory response (GO:0006954), negative regulation of tumor necrosis factor-mediated signaling pathway (GO:0010804), protein destabilization (GO:0031648), negative regulation of lipopolysaccharide-mediated signaling pathway (GO:0031665), negative regulation of interleukin-1 beta production (GO:0032691), positive regulation of interleukin-1 beta production (GO:0032731), regulation of apoptotic process (GO:0042981), negative regulation of canonical NF-kappaB signal transduction (GO:0043124), positive regulation of inflammatory response (GO:0050729), defense response to virus (GO:0051607), intrinsic apoptotic signaling pathway (GO:0097193), self proteolysis (GO:0097264), antiviral innate immune response (GO:0140374), CARD8 inflammasome complex assembly (GO:0140633), negative regulation of NLRP3 inflammasome complex assembly (GO:1900226), immune system process (GO:0002376), proteolysis (GO:0006508), programmed cell death (GO:0012501), innate immune response (GO:0045087), protein maturation (GO:0051604), pyroptotic cell death (GO:0141201)

GO Molecular Function (9): peptidase activity (GO:0008233), NACHT domain binding (GO:0032089), pattern recognition receptor activity (GO:0038187), protein homodimerization activity (GO:0042803), CARD domain binding (GO:0050700), cysteine-type endopeptidase activator activity (GO:0140608), molecular condensate scaffold activity (GO:0140693), protein binding (GO:0005515), hydrolase activity (GO:0016787)

GO Cellular Component (8): nucleus (GO:0005634), nucleoplasm (GO:0005654), cytoplasm (GO:0005737), cytosol (GO:0005829), protein-containing complex (GO:0032991), NLRP3 inflammasome complex (GO:0072559), CARD8 inflammasome complex (GO:0140634), canonical inflammasome complex (GO:0061702)

Reactome top-level categories

Rollup of top-6 pathways:

Category	Pathways
Cytochrome c-mediated apoptotic response	1
Formation of apoptosome	1
Programmed Cell Death	1
Apoptosis	1
Apoptotic factor-mediated response	1
Intrinsic Pathway for Apoptosis	1

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
cellular anatomical structure	3
defense response	2
interleukin-1 beta production	2
regulation of interleukin-1 beta production	2
protein domain specific binding	2
canonical inflammasome complex	2
activation of immune response	1
positive regulation of innate immune response	1
negative regulation of cytokine-mediated signaling pathway	1
regulation of tumor necrosis factor-mediated signaling pathway	1
tumor necrosis factor-mediated signaling pathway	1
regulation of protein stability	1
negative regulation of response to biotic stimulus	1
negative regulation of signal transduction	1
lipopolysaccharide-mediated signaling pathway	1
regulation of lipopolysaccharide-mediated signaling pathway	1
negative regulation of response to external stimulus	1
negative regulation of interleukin-1 production	1
positive regulation of interleukin-1 production	1
apoptotic process	1
regulation of programmed cell death	1
canonical NF-kappaB signal transduction	1
regulation of canonical NF-kappaB signal transduction	1
negative regulation of intracellular signal transduction	1
inflammatory response	1
positive regulation of defense response	1
positive regulation of response to external stimulus	1
regulation of inflammatory response	1
response to virus	1
intracellular signal transduction	1
apoptotic signaling pathway	1
proteolysis	1
innate immune response	1
defense response to virus	1
canonical inflammasome complex assembly	1
negative regulation of protein-containing complex assembly	1
NLRP3 inflammasome complex assembly	1
negative regulation of inflammasome-mediated signaling pathway	1
regulation of NLRP3 inflammasome complex assembly	1
biological_process	1

Protein interactions and networks

STRING

776 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
CARD8	NLRP3	Q96P20	976
CARD8	CASP1	P29466	973
CARD8	PYCARD	Q9ULZ3	967
CARD8	FHL2	Q14192	915
CARD8	CASP5	P51878	900
CARD8	NLRP2	Q9NX02	880
CARD8	MEFV	O15553	845
CARD8	AIM2	O14862	809
CARD8	NLRC4	Q9NPP4	799
CARD8	CARD16	Q5EG05	776
CARD8	DPP9	Q86TI2	737
CARD8	CARD6	Q9BX69	728
CARD8	MINDY3	Q9H8M7	713
CARD8	CASP9	P55211	700
CARD8	IL18	Q14116	694

IntAct

17 interactions, top by confidence:

A	B	Type	Score
CARD8	CARD8	psi-mi:“MI:0915”(physical association)	0.510
CARD8	FHL2	psi-mi:“MI:0915”(physical association)	0.510
FHL2	CARD8	psi-mi:“MI:0915”(physical association)	0.510
NLRP3	CARD8	psi-mi:“MI:0915”(physical association)	0.400
CASP9	CARD8	psi-mi:“MI:0915”(physical association)	0.400
IKBKG	CARD8	psi-mi:“MI:0915”(physical association)	0.400
LHX6	KDM5C	psi-mi:“MI:0914”(association)	0.350
CARD8	HDAC3	psi-mi:“MI:0914”(association)	0.350
rep	VWA8	psi-mi:“MI:0914”(association)	0.350
CARD8	FLNA	psi-mi:“MI:0914”(association)	0.350
SFN	BLTP3B	psi-mi:“MI:2364”(proximity)	0.270
lpd	CARD8	psi-mi:“MI:0915”(physical association)	0.000
tuf	CARD8	psi-mi:“MI:0915”(physical association)	0.000
	CARD8	psi-mi:“MI:0915”(physical association)	0.000

BioGRID (112): FLNA (Affinity Capture-MS), FLNC (Affinity Capture-MS), CSRP2BP (Affinity Capture-MS), DMAP1 (Affinity Capture-MS), BRCA2 (Affinity Capture-MS), YEATS2 (Affinity Capture-MS), TAB1 (Affinity Capture-MS), BCOR (Affinity Capture-MS), CEP192 (Affinity Capture-MS), NFRKB (Affinity Capture-MS), SRCAP (Affinity Capture-MS), ZZZ3 (Affinity Capture-MS), ZNF318 (Affinity Capture-MS), LRIF1 (Affinity Capture-MS), MGA (Affinity Capture-MS)

ESM2 similar proteins: A6H5X4, D0QMC3, D2HHP1, G3HKI1, O14862, O35368, P0DOV1, P0DOV2, P41218, P62597, Q05CL8, Q13287, Q13426, Q14159, Q16666, Q4G0J3, Q4R627, Q4R7Q1, Q504N7, Q5I0E2, Q5RAV7, Q5RCV3, Q5RCZ8, Q5RD14, Q60953, Q66JT0, Q68D51, Q6IEE8, Q6K0P9, Q6ZMT9, Q8BGX7, Q8BV49, Q8C0V1, Q8C267, Q8CGE8, Q8IYM2, Q8NAT2, Q8NDB2, Q8NE18, Q8SPH9

Diamond homologs: A1Z198, A6QLE5, A8Y3R9, B0FPE9, D4A523, D9I2F9, D9I2G1, D9I2G3, D9I2G4, D9I2H0, E9Q5R7, P10775, P13489, P29315, P59044, P59046, P59047, Q0GKD5, Q2LKU9, Q2LKV2, Q2LKV5, Q2LKW6, Q5RAV7, Q63035, Q6B966, Q86W24, Q86W25, Q86W26, Q8CCN1, Q8HXK9, Q8HZP9, Q8R4B8, Q91VI7, Q91WS2, Q96P20, Q9C000, Q9EPB4, Q9I9N6, Q9ULZ3, Q9Y2G2

SIGNOR signaling

1 interactions.

A	Effect	B	Mechanism
CARD8	down-regulates	CASP9	binding

Disease & clinical

Clinical variants and AI predictions

ClinVar

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

Classification	Count (floor)
Pathogenic	0
Likely pathogenic	1
Uncertain significance	296
Likely benign	158
Benign	26

Top pathogenic / likely-pathogenic (1)

Variant ID	HGVS	Classification
4687999	NM_001184900.3(CARD8):c.180dup (p.Gln61fs)	Likely pathogenic

SpliceAI

3027 predictions. Top by Δscore:

Variant	Effect	Δscore
19:48194638:T:TA	acceptor_gain	1.0000
19:48194644:C:A	acceptor_gain	1.0000
19:48194651:A:AG	acceptor_gain	1.0000
19:48194651:AG:A	acceptor_gain	1.0000
19:48194651:AGGT:A	acceptor_gain	1.0000
19:48194652:G:GT	acceptor_gain	1.0000
19:48194652:GG:G	acceptor_gain	1.0000
19:48194652:GGT:G	acceptor_gain	1.0000
19:48194652:GGTG:G	acceptor_gain	1.0000
19:48194652:GGTGA:G	acceptor_gain	1.0000
19:48211976:C:CC	acceptor_gain	1.0000
19:48215382:CCA:C	acceptor_gain	1.0000
19:48215383:CA:C	acceptor_gain	1.0000
19:48215383:CAC:C	acceptor_gain	1.0000
19:48215385:C:CC	acceptor_gain	1.0000
19:48221725:CTAA:C	donor_loss	1.0000
19:48221726:TAAC:T	donor_loss	1.0000
19:48221727:AAC:A	donor_loss	1.0000
19:48221728:A:AG	donor_loss	1.0000
19:48221729:CCTTG:C	donor_loss	1.0000
19:48221857:T:C	acceptor_loss	1.0000
19:48230433:CTTA:C	donor_loss	1.0000
19:48230436:A:C	donor_loss	1.0000
19:48230437:C:T	donor_loss	1.0000
19:48230701:C:CC	acceptor_gain	1.0000
19:48230908:C:T	acceptor_gain	1.0000
19:48232511:CATGT:C	acceptor_gain	1.0000
19:48234397:TCTTA:T	donor_loss	1.0000
19:48234398:CTTA:C	donor_loss	1.0000
19:48234399:TTACC:T	donor_loss	1.0000

AlphaMissense

0 scored. Top likely-pathogenic:

dbSNP variants (sampled 300 via entrez): RS1000004962 (19:48182503 G>A), RS1000013643 (19:48183367 T>C), RS1000040692 (19:48206876 T>C), RS1000067163 (19:48217153 G>A), RS1000071585 (19:48183489 G>A,C), RS1000098382 (19:48253631 G>A,T), RS1000142189 (19:48218481 C>T), RS1000175723 (19:48188458 G>T), RS1000250749 (19:48222910 T>G), RS1000313140 (19:48222264 T>C,G), RS1000371141 (19:48247161 G>A,T), RS1000411636 (19:48194274 C>T), RS1000455694 (19:48228295 T>C), RS1000508663 (19:48221497 A>G), RS1000523313 (19:48216451 G>A,C)

Disease associations

OMIM: gene MIM:609051 | disease phenotypes: MIM:619079

GenCC curated gene-disease

Disease	Classification	Inheritance
inflammatory bowel disease 30	Limited	Unknown

Mondo (1): inflammatory bowel disease 30 (MONDO:0033643)

Orphanet (0):

HPO phenotypes

13 total (13 of 13 shown, HPO-id order):

HPO	Term
HP:0000006	Autosomal dominant inheritance
HP:0002013	Vomiting
HP:0002027	Abdominal pain
HP:0002028	Chronic diarrhea
HP:0002242	Abnormal intestine morphology
HP:0002243	Protein-losing enteropathy
HP:0005263	Gastritis
HP:0020002	Ileal stricture
HP:0025085	Bloody diarrhea
HP:0032564	Ileitis
HP:0033117	Duodenitis
HP:0033256	Pancolitis
HP:0100633	Esophagitis

GWAS associations

0 associations (top):

Drugs & pharmacology

Drug and pharmacology data

Is drug target: no

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

PharmGKB variants

1 variants.

Variant	Genes	Level	Score	#Clin annots	Drugs
rs2043211	CARD8		0.00	0

CTD chemical–gene interactions

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

Chemical	Actions (top 5)	PubMed papers
Valproic Acid	affects cotreatment, decreases expression, affects expression	9
Phenylmercuric Acetate	affects cotreatment, decreases expression	2
triphenyl phosphate	affects expression	1
terbufos	decreases methylation	1
tris(1,3-dichloro-2-propyl)phosphate	decreases expression	1
sodium arsenite	decreases expression	1
butyraldehyde	decreases expression	1
manganese chloride	increases abundance, increases expression	1
CGP 52608	affects binding, increases reaction	1
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide	affects cotreatment, decreases expression	1
dorsomorphin	affects cotreatment, decreases expression	1
NSC 689534	affects binding, increases expression	1
Temozolomide	increases expression	1
Sunitinib	decreases expression	1
Zoledronic Acid	increases expression	1
Acetaminophen	decreases expression	1
Asbestos	affects response to substance	1
Vehicle Emissions	affects expression, increases abundance	1
Benzo(a)pyrene	decreases methylation	1
Cannabidiol	decreases expression	1
Copper	affects binding, increases expression	1
Coumestrol	increases expression	1
Dietary Carbohydrates	increases expression	1
Doxorubicin	decreases expression	1
Fonofos	decreases methylation	1
Manganese	increases expression, increases abundance	1
Menthol	increases expression	1
Methyl Methanesulfonate	increases expression	1
Paraquat	decreases expression	1
Parathion	decreases methylation	1

Cellosaurus cell lines

2 cell lines: 2 cancer cell line

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

Cellosaurus	Name	Category	Sex
CVCL_E0WG	Ubigene Jurkat, Clone E6-1 CARD8 KO	Cancer cell line	Male
CVCL_E1CR	Ubigene THP-1 CARD8 KO	Cancer cell line	Male

Clinical trials (associated diseases)

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