{"gene":"CRACR2A","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":2010,"finding":"CRACR2A (EFCAB4B) directly interacts with both Orai1 and STIM1, forming a ternary complex that dissociates at elevated Ca2+ concentrations. CRACR2A is required for co-clustering of Orai1 and STIM1 upon store depletion, and an EF-hand mutant of CRACR2A that cannot sense Ca2+ constitutively enhanced STIM1 clustering, elevated cytoplasmic Ca2+, and induced cell death.","method":"Affinity protein purification, siRNA knockdown, EF-hand mutagenesis, co-immunoprecipitation, confocal imaging of Orai1/STIM1 clusters","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP establishing ternary complex, siRNA knockdown with defined phenotype, and mutagenesis all in one study with multiple orthogonal methods","pmids":["20418871"],"is_preprint":false},{"year":2016,"finding":"The CRACR2A locus encodes a lymphocyte-specific large Rab GTPase isoform containing EF-hand motifs, a proline-rich domain (PRD), and a Rab GTPase domain with an unconventional prenylation site. This isoform localizes to vesicles near the Golgi (dependent on GTP binding and prenylation), which translocate to the immunological synapse upon TCR stimulation to activate Ca2+/NFAT and JNK signaling pathways. The interaction between the PRD of CRACR2A and the GEF Vav1 is required for accumulation of these vesicles at the immunological synapse.","method":"Gene silencing, knockout mice, domain mutagenesis (GTP-binding and prenylation mutants), co-immunoprecipitation of CRACR2A-PRD with Vav1, live-cell imaging of vesicle translocation, Ca2+ and JNK signaling assays","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KO mice, domain mutants, Co-IP, live imaging, signaling readouts) establishing pathway position and molecular interactions","pmids":["27016526"],"is_preprint":false},{"year":2019,"finding":"CRACR2A acts as a calcium-activated dynein adaptor: its coiled-coil domain joins dynein and dynactin into a motile complex, and its EF-hand domain activates this motility in a calcium-dependent manner in vitro. In Jurkat T cells, elevation of intracellular calcium activates CRACR2A-mediated dynein transport, and TCR activation induces formation of CRACR2A puncta at the plasma membrane that associate with the actin cortex and then travel along microtubules in an endocytic process.","method":"In vitro dynein motility reconstitution assay, intracellular Ca2+ manipulation in Jurkat T cells, live-cell imaging of CRACR2A puncta, microtubule and actin co-localization","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of Ca2+-activated dynein motility plus live-cell imaging demonstrating endocytic transport, single lab but multiple orthogonal methods","pmids":["30814157"],"is_preprint":false},{"year":2019,"finding":"Rab46 (CRACR2A long isoform) in endothelial cells integrates histamine (H1 receptor/G protein) and Ca2+ signals to regulate selective Weibel-Palade body (WPB) trafficking: GTP-bound Rab46 on a P-selectin-negative WPB subset mediates dynein-dependent retrograde transport to the microtubule organizing center (MTOC) upon histamine stimulation, and subsequently Ca2+ binding to Rab46's EF-hand triggers dispersal of MTOC-clustered WPBs, thereby diverting non-inflammatory cargo away from the plasma membrane.","method":"Rab46 localization by imaging, GTP/GDP-locked Rab46 mutants, dynein inhibition, intracellular Ca2+ measurement, selective cargo release assay (P-selectin vs. non-inflammatory cargo)","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — nucleotide-locked mutants combined with dynein inhibition and Ca2+ imaging provide multiple orthogonal lines of evidence, single lab","pmids":["31092558"],"is_preprint":false},{"year":2014,"finding":"Endothelial cells express a long variant of CRACR2A (CRACR2A-L / Rab46) that is approximately twice the molecular mass of the short form and contains an additional C-terminal Rab GTPase domain; siRNA against CRACR2A in endothelial cells depletes this long form but has no effect on CRAC channel function, and CRACR2A-L makes a positive contribution to endothelial tube formation.","method":"siRNA knockdown, Western blot, full-length cDNA cloning, sequence analysis, endothelial tube formation assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — identifies the long isoform by cloning and links it to tube formation via KD, but functional mechanism is not fully resolved; negative finding on CRAC channel role is also established","pmids":["25475730"],"is_preprint":false},{"year":2018,"finding":"CRACR2A promotes Th1 differentiation and effector function of Th17 cells downstream of TCR signaling: conditional T cell-specific deletion of CRACR2A in mice reduced Th1 responses to LCMV infection and conferred resistance to EAE, with impaired Ca2+/NFAT and JNK/AP1 pathway activation due to deficient TCR signal transmission.","method":"Conditional knockout mice (T cell-specific Cre), viral infection model, EAE model, transcript analysis, Ca2+ and JNK signaling assays","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean conditional KO with multiple in vivo disease models and defined signaling readouts, corroborated by prior mechanistic work from same group","pmids":["29987160"],"is_preprint":false},{"year":2021,"finding":"Biallelic loss-of-function mutations in CRACR2A cause late-onset combined immunodeficiency in humans. Patient T cells show reduced CRACR2A protein, dampened SOCE (Ca2+ entry), and reduced JNK phosphorylation. Reconstitution experiments in CRACR2A-deleted T cells showed that the E278D allele specifically impairs SOCE/cytokine production (not JNK), while the R144G/E300* truncation impairs both JNK phosphorylation and SOCE, establishing allele-specific separation of CRACR2A's two signaling functions.","method":"Patient allele characterization, CRACR2A KO T cell reconstitution with individual mutant alleles, SOCE measurement (Ca2+ imaging), JNK phosphorylation assay, cytokine production assay","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 / Strong — allele-specific reconstitution in KO T cells with multiple orthogonal readouts (SOCE, JNK, cytokines) rigorously separates two molecular functions","pmids":["34908525"],"is_preprint":false},{"year":2021,"finding":"Affinity purification coupled to LC-MS/MS identified dynein and Na+/K+ ATPase subunit alpha 1 (ATP1α1) as validated binding partners (effectors) of GTP-bound Rab46 (CRACR2A) in endothelial cells, confirmed by biochemical and imaging approaches.","method":"Affinity purification with constitutively active vs. inactive GFP-Rab46, LC-MS/MS, biochemical validation, imaging","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — affinity purification with nucleotide-locked mutants plus biochemical validation, but limited mechanistic follow-up on ATP1α1 interaction","pmids":["33603063"],"is_preprint":false},{"year":2024,"finding":"In neutrophils, CRACR2A rapidly associates with STIM1 after agonist stimulation and facilitates Ca2+ mobilization, which increases the ligand-binding function of β2 integrin and promotes neutrophil adhesion, crawling, and transmigration. A palmitoylated 20-mer peptide from the coiled-coil region of CRACR2A blocks STIM1-CRACR2A interaction, inhibiting Ca2+ mobilization and β2 integrin activation.","method":"Myeloid-specific Cracr2a conditional KO mice, CRISPR/Cas9 KO in dHL-60 cells, CRACR2A rescue overexpression, immunoprecipitation of CRACR2A-STIM1 complex, cytosolic Ca2+ mobilization assay, flow cytometry for integrin activation, intravital microscopy, blocking peptide experiments","journal":"Circulation","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO mice plus CRISPR KO plus rescue plus peptide inhibitor plus multiple functional readouts (Co-IP, Ca2+, integrin activation, intravital microscopy)","pmids":["39601147"],"is_preprint":false},{"year":2025,"finding":"Ca2+ released specifically from TPC2 (two-pore channel 2) endolysosomal channels is required for the EF-hand-dependent detachment of Rab46 (CRACR2A) from the MTOC, thereby enabling secretion of Ang2-positive Weibel-Palade bodies in endothelial cells. Pharmacological TPC2 inhibition increased Rab46 clustering at the MTOC and decreased Ang2 secretion, while a TPC2 agonist had the opposite effect.","method":"Ca2+ imaging, high-resolution light microscopy, pharmacological inhibition/activation of TPC2 (Ned19, tetrandrine, TPC2-A1-N), Rab46 localization at MTOC, Ang2 secretion assay","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological approach with imaging and secretion readouts; preprint, single lab, no genetic confirmation of TPC2 specificity","pmids":["bio_10.1101_2025.03.03.641167"],"is_preprint":true}],"current_model":"CRACR2A (EFCAB4B) is a multifunctional Ca2+-sensing protein expressed as a short isoform (EF-hand CRAC channel regulator) and a long isoform (Rab46, a large Rab GTPase) that: (1) forms a Ca2+-dissociable ternary complex with Orai1 and STIM1 to stabilize CRAC channel clusters and promote store-operated Ca2+ entry; (2) acts as a lymphocyte-specific vesicle-associated large Rab GTPase whose PRD-Vav1 interaction drives vesicle translocation to the immunological synapse to activate Ca2+/NFAT and JNK/AP1 TCR signaling; (3) functions as a calcium-activated dynein adaptor, coupling EF-hand Ca2+ sensing to dynein-mediated retrograde endocytic transport; (4) in endothelial cells, integrates G protein and Ca2+ signals (including TPC2-derived Ca2+) to control selective Weibel-Palade body trafficking via dynein; and (5) in neutrophils, associates with STIM1 after agonist stimulation to facilitate Ca2+ mobilization and β2 integrin activation, promoting neutrophil recruitment."},"narrative":{"mechanistic_narrative":"CRACR2A (EFCAB4B) is a Ca2+-sensing protein, expressed as a short EF-hand isoform and a long large-Rab-GTPase isoform (Rab46), that couples calcium signaling to two distinct effector outputs: store-operated Ca2+ entry and dynein-based vesicle transport [PMID:20418871, PMID:30814157, PMID:31092558]. The short isoform directly binds both Orai1 and STIM1 to form a Ca2+-dissociable ternary complex that stabilizes Orai1/STIM1 co-clustering and supports CRAC channel function; constitutive (Ca2+-insensitive) EF-hand mutants drive aberrant STIM1 clustering, Ca2+ overload, and cell death [PMID:20418871]. In lymphocytes, a large isoform bearing EF-hand motifs, a proline-rich domain (PRD), and a prenylated Rab GTPase domain localizes to Golgi-proximal vesicles whose PRD–Vav1 interaction drives translocation to the immunological synapse, transmitting TCR signals through the Ca2+/NFAT and JNK/AP1 pathways to support Th1 and Th17 effector responses [PMID:27016526, PMID:29987160]. Mechanistically, CRACR2A functions as a calcium-activated dynein adaptor: its coiled-coil domain assembles dynein and dynactin into a motile complex and its EF-hand domain confers Ca2+-dependent activation of motility, linking Ca2+ rises to retrograde endocytic transport [PMID:30814157]. In endothelial cells the Rab46 isoform integrates histamine-driven G protein and Ca2+ signals to selectively route a P-selectin-negative Weibel-Palade body subset to the MTOC via dynein, with EF-hand Ca2+ binding triggering subsequent dispersal and selective cargo release [PMID:31092558, PMID:bio_10.1101_2025.03.03.641167]. In neutrophils CRACR2A associates with STIM1 after agonist stimulation to drive Ca2+ mobilization and β2 integrin activation, promoting recruitment [PMID:39601147]. Biallelic loss-of-function mutations in CRACR2A cause late-onset combined immunodeficiency, and allele-specific reconstitution separates its SOCE function from its JNK-activating function [PMID:34908525].","teleology":[{"year":2010,"claim":"Established CRACR2A as a Ca2+-sensing regulator of CRAC channels by showing it bridges Orai1 and STIM1 in a complex that disassembles when Ca2+ rises.","evidence":"Affinity purification, reciprocal Co-IP, siRNA knockdown, and EF-hand mutagenesis with confocal cluster imaging","pmids":["20418871"],"confidence":"High","gaps":["Structural basis of the ternary complex not resolved","Did not address the existence of a second large isoform"]},{"year":2014,"claim":"Revealed that the CRACR2A locus produces a long endothelial isoform (Rab46) with an additional Rab GTPase domain that is functionally distinct from the CRAC-channel-regulating short form.","evidence":"cDNA cloning, Western blot, siRNA knockdown, and endothelial tube formation assay","pmids":["25475730"],"confidence":"Medium","gaps":["Mechanism of contribution to tube formation unresolved","GTPase domain function not yet characterized"]},{"year":2016,"claim":"Defined the lymphocyte large isoform as a vesicle-associated Rab GTPase that uses a PRD-Vav1 interaction to deliver TCR signaling to the immunological synapse, positioning CRACR2A in NFAT and JNK pathways.","evidence":"Knockout mice, GTP-binding/prenylation mutants, CRACR2A-PRD/Vav1 Co-IP, live-cell vesicle imaging, and signaling assays","pmids":["27016526"],"confidence":"High","gaps":["Identity of vesicle cargo at the synapse not defined","How Vav1 GEF activity feeds back on CRACR2A unclear"]},{"year":2018,"claim":"Demonstrated the physiological consequence of CRACR2A loss in T cells, linking it to Th1/Th17 effector function and in vivo immune responses.","evidence":"T cell-specific conditional knockout mice in LCMV and EAE models with Ca2+/NFAT and JNK readouts","pmids":["29987160"],"confidence":"High","gaps":["Does not separate the SOCE versus JNK contributions to disease phenotypes"]},{"year":2019,"claim":"Established the core molecular mechanism: CRACR2A is a calcium-activated dynein adaptor whose coiled-coil assembles dynein-dynactin and whose EF-hand confers Ca2+-dependent motility, mediating endocytic transport.","evidence":"In vitro dynein motility reconstitution plus Ca2+ manipulation and live-cell puncta imaging in Jurkat T cells","pmids":["30814157"],"confidence":"High","gaps":["Cargo of the endocytic transport process not identified","Single lab"]},{"year":2019,"claim":"Showed that the Rab46 isoform performs selective organelle sorting in endothelium, using dynein for MTOC retrograde transport and EF-hand Ca2+ sensing to disperse a specific WPB subset.","evidence":"Nucleotide-locked Rab46 mutants, dynein inhibition, Ca2+ imaging, and selective cargo release assays","pmids":["31092558"],"confidence":"High","gaps":["Source of the dispersal-triggering Ca2+ not yet defined","Single lab"]},{"year":2021,"claim":"Identified the GTP-bound Rab46 effector proteome, confirming dynein and adding Na+/K+ ATPase subunit ATP1α1 as direct partners.","evidence":"Affinity purification of constitutively active vs inactive GFP-Rab46 with LC-MS/MS and biochemical/imaging validation","pmids":["33603063"],"confidence":"Medium","gaps":["Functional role of the ATP1α1 interaction not pursued","No mechanistic follow-up"]},{"year":2021,"claim":"Proved CRACR2A is disease-relevant in humans and genetically dissected its two signaling functions via allele-specific reconstitution.","evidence":"Patient allele characterization and CRACR2A-KO T cell reconstitution with SOCE, JNK, and cytokine readouts","pmids":["34908525"],"confidence":"High","gaps":["Structural basis for the E278D SOCE-specific defect not defined","Spectrum of clinical phenotypes incompletely mapped"]},{"year":2024,"claim":"Extended CRACR2A function to innate immunity, showing a STIM1-coupled Ca2+ mobilization role that activates β2 integrin and drives neutrophil recruitment, and provided a peptide inhibitor.","evidence":"Myeloid-specific KO mice, CRISPR KO in dHL-60 cells, rescue, CRACR2A-STIM1 Co-IP, Ca2+ and integrin assays, intravital microscopy, and a coiled-coil blocking peptide","pmids":["39601147"],"confidence":"High","gaps":["Whether neutrophil function uses the short or large isoform not fully resolved","Relationship to dynein-adaptor activity in neutrophils unaddressed"]},{"year":2025,"claim":"Pinpointed TPC2 endolysosomal Ca2+ as the specific signal driving EF-hand-dependent Rab46 detachment from the MTOC to enable Ang2-positive WPB secretion.","evidence":"Ca2+ imaging, high-resolution microscopy, pharmacological TPC2 inhibition/activation, and Ang2 secretion assays (preprint)","pmids":["bio_10.1101_2025.03.03.641167"],"confidence":"Medium","gaps":["No genetic confirmation of TPC2 specificity","Single lab, preprint"]},{"year":null,"claim":"How the short CRAC-regulatory isoform and the large dynein-adaptor/Rab isoform are differentially deployed across cell types, and whether their functions intersect within a single cell, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified structural model linking EF-hand Ca2+ sensing, coiled-coil dynein assembly, and Rab GTPase activity","Isoform-specific upstream regulation not mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140299","term_label":"molecular sensor activity","supporting_discovery_ids":[0,2,3]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2]},{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[1,3,7]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,8]}],"localization":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[1,3]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[3,9]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,2]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,5,6,8]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,3]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[2,3]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3,9]}],"complexes":["Orai1-STIM1-CRACR2A ternary CRAC complex","dynein-dynactin motile complex"],"partners":["ORAI1","STIM1","VAV1","ATP1A1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BSW2","full_name":"EF-hand calcium-binding domain-containing protein 4B","aliases":["Calcium release-activated calcium channel regulator 2A","CRAC channel regulator 2A","Calcium release-activated channel regulator 2A","Ras-related protein Rab-46"],"length_aa":731,"mass_kda":83.2,"function":"Ca(2+)-binding protein that plays a key role in store-operated Ca(2+) entry (SOCE) in T-cells by regulating CRAC channel activation. Acts as a cytoplasmic calcium-sensor that facilitates the clustering of ORAI1 and STIM1 at the junctional regions between the plasma membrane and the endoplasmic reticulum upon low Ca(2+) concentration. It thereby regulates CRAC channel activation, including translocation and clustering of ORAI1 and STIM1. Upon increase of cytoplasmic Ca(2+) resulting from opening of CRAC channels, dissociates from ORAI1 and STIM1, thereby destabilizing the ORAI1-STIM1 complex Rab GTPase that mediates the trafficking of Weibel-Palade bodies (WPBs) to microtubule organizing center (MTOC) in endothelial cells in response to acute inflammatory stimuli (PubMed:31092558). During histamine (but not thrombin) stimulation of endothelial cells, the dynein-bound form induces retrograde transport of a subset of WPBs along microtubules to the MTOC in a Ca(2+)-independent manner and its GTPase activity is essential for this function (PubMed:31092558). Ca(2+)-regulated dynein adapter protein that activates dynein-mediated transport and dynein-dynactin motility on microtubules and regulates endosomal trafficking of CD47 (PubMed:30814157). Acts as an intracellular signaling module bridging two important T-cell receptor (TCR) signaling pathways, Ca(2+)-NFAT and JNK, to affect T-cell activation (PubMed:27016526). In resting T-cells, is predominantly localized near TGN network in a GTP-bound form, upon TCR stimulation, localizes at the immunological synapse via interaction with VAV1 to activate downstream Ca(2+)-NFAT and JNK signaling pathways (PubMed:27016526). Plays a role in T-helper 1 (Th1) cell differentiation and T-helper 17 (Th17) cell effector function (PubMed:29987160). Plays a role in store-operated Ca(2+) entry (SOCE) in T-cells by regulating CRAC channel activation (PubMed:27016526)","subcellular_location":"Cytoplasm; Cytoplasm, cytoskeleton, microtubule organizing center; Cell membrane; Golgi apparatus membrane; Golgi apparatus, trans-Golgi network membrane; Vesicle","url":"https://www.uniprot.org/uniprotkb/Q9BSW2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CRACR2A","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CRACR2A","total_profiled":1310},"omim":[{"mim_id":"614178","title":"CALCIUM RELEASE-ACTIVATED CALCIUM CHANNEL REGULATOR 2A; CRACR2A","url":"https://www.omim.org/entry/614178"},{"mim_id":"614177","title":"EF-HAND CALCIUM-BINDING DOMAIN-CONTAINING PROTEIN 4A; EFCAB4A","url":"https://www.omim.org/entry/614177"},{"mim_id":"610277","title":"ORAI CALCIUM RELEASE-ACTIVATED CALCIUM MODULATOR 1; ORAI1","url":"https://www.omim.org/entry/610277"},{"mim_id":"605921","title":"STROMAL INTERACTION MOLECULE 1; STIM1","url":"https://www.omim.org/entry/605921"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":16.0},{"tissue":"intestine","ntpm":12.7},{"tissue":"salivary gland","ntpm":36.4}],"url":"https://www.proteinatlas.org/search/CRACR2A"},"hgnc":{"alias_symbol":["MGC4266","RAB46"],"prev_symbol":["EFCAB4B"]},"alphafold":{"accession":"Q9BSW2","domains":[{"cath_id":"1.10.238.10","chopping":"48-120","consensus_level":"high","plddt":77.2844,"start":48,"end":120},{"cath_id":"3.40.50.300","chopping":"545-717","consensus_level":"high","plddt":88.7382,"start":545,"end":717}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BSW2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BSW2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BSW2-F1-predicted_aligned_error_v6.png","plddt_mean":69.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CRACR2A","jax_strain_url":"https://www.jax.org/strain/search?query=CRACR2A"},"sequence":{"accession":"Q9BSW2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BSW2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BSW2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BSW2"}},"corpus_meta":[{"pmid":"20418871","id":"PMC_20418871","title":"A novel EF-hand protein, CRACR2A, is a cytosolic Ca2+ sensor that stabilizes CRAC channels in T cells.","date":"2010","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/20418871","citation_count":196,"is_preprint":false},{"pmid":"30814157","id":"PMC_30814157","title":"CRACR2a is a calcium-activated dynein adaptor protein that regulates endocytic traffic.","date":"2019","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/30814157","citation_count":44,"is_preprint":false},{"pmid":"27016526","id":"PMC_27016526","title":"A large Rab GTPase encoded by CRACR2A is a component of subsynaptic vesicles that transmit T cell activation signals.","date":"2016","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/27016526","citation_count":32,"is_preprint":false},{"pmid":"31092558","id":"PMC_31092558","title":"Rab46 integrates Ca2+ and histamine signaling to regulate selective cargo release from Weibel-Palade 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immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38567292","citation_count":2,"is_preprint":false},{"pmid":"40034259","id":"PMC_40034259","title":"RNA-seq analysis reveals transcriptome changes in livers from Efcab4b knockout mice.","date":"2025","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/40034259","citation_count":1,"is_preprint":false},{"pmid":"35151051","id":"PMC_35151051","title":"CRACR2A associated immunodeficiency - the supporting factor takes center stage.","date":"2022","source":"Cell calcium","url":"https://pubmed.ncbi.nlm.nih.gov/35151051","citation_count":1,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.03.03.641167","title":"Two-pore channel protein 2-mediated calcium release promotes angiopoietin 2 secretion by regulating Rab46-dependent Weibel-Palade body trafficking","date":"2025-03-03","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.03.641167","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9765,"output_tokens":3290,"usd":0.039323,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10731,"output_tokens":3839,"usd":0.074815,"stage2_stop_reason":"end_turn"},"total_usd":0.114138,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2010,\n      \"finding\": \"CRACR2A (EFCAB4B) directly interacts with both Orai1 and STIM1, forming a ternary complex that dissociates at elevated Ca2+ concentrations. CRACR2A is required for co-clustering of Orai1 and STIM1 upon store depletion, and an EF-hand mutant of CRACR2A that cannot sense Ca2+ constitutively enhanced STIM1 clustering, elevated cytoplasmic Ca2+, and induced cell death.\",\n      \"method\": \"Affinity protein purification, siRNA knockdown, EF-hand mutagenesis, co-immunoprecipitation, confocal imaging of Orai1/STIM1 clusters\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP establishing ternary complex, siRNA knockdown with defined phenotype, and mutagenesis all in one study with multiple orthogonal methods\",\n      \"pmids\": [\"20418871\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The CRACR2A locus encodes a lymphocyte-specific large Rab GTPase isoform containing EF-hand motifs, a proline-rich domain (PRD), and a Rab GTPase domain with an unconventional prenylation site. This isoform localizes to vesicles near the Golgi (dependent on GTP binding and prenylation), which translocate to the immunological synapse upon TCR stimulation to activate Ca2+/NFAT and JNK signaling pathways. The interaction between the PRD of CRACR2A and the GEF Vav1 is required for accumulation of these vesicles at the immunological synapse.\",\n      \"method\": \"Gene silencing, knockout mice, domain mutagenesis (GTP-binding and prenylation mutants), co-immunoprecipitation of CRACR2A-PRD with Vav1, live-cell imaging of vesicle translocation, Ca2+ and JNK signaling assays\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KO mice, domain mutants, Co-IP, live imaging, signaling readouts) establishing pathway position and molecular interactions\",\n      \"pmids\": [\"27016526\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CRACR2A acts as a calcium-activated dynein adaptor: its coiled-coil domain joins dynein and dynactin into a motile complex, and its EF-hand domain activates this motility in a calcium-dependent manner in vitro. In Jurkat T cells, elevation of intracellular calcium activates CRACR2A-mediated dynein transport, and TCR activation induces formation of CRACR2A puncta at the plasma membrane that associate with the actin cortex and then travel along microtubules in an endocytic process.\",\n      \"method\": \"In vitro dynein motility reconstitution assay, intracellular Ca2+ manipulation in Jurkat T cells, live-cell imaging of CRACR2A puncta, microtubule and actin co-localization\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of Ca2+-activated dynein motility plus live-cell imaging demonstrating endocytic transport, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"30814157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Rab46 (CRACR2A long isoform) in endothelial cells integrates histamine (H1 receptor/G protein) and Ca2+ signals to regulate selective Weibel-Palade body (WPB) trafficking: GTP-bound Rab46 on a P-selectin-negative WPB subset mediates dynein-dependent retrograde transport to the microtubule organizing center (MTOC) upon histamine stimulation, and subsequently Ca2+ binding to Rab46's EF-hand triggers dispersal of MTOC-clustered WPBs, thereby diverting non-inflammatory cargo away from the plasma membrane.\",\n      \"method\": \"Rab46 localization by imaging, GTP/GDP-locked Rab46 mutants, dynein inhibition, intracellular Ca2+ measurement, selective cargo release assay (P-selectin vs. non-inflammatory cargo)\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — nucleotide-locked mutants combined with dynein inhibition and Ca2+ imaging provide multiple orthogonal lines of evidence, single lab\",\n      \"pmids\": [\"31092558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Endothelial cells express a long variant of CRACR2A (CRACR2A-L / Rab46) that is approximately twice the molecular mass of the short form and contains an additional C-terminal Rab GTPase domain; siRNA against CRACR2A in endothelial cells depletes this long form but has no effect on CRAC channel function, and CRACR2A-L makes a positive contribution to endothelial tube formation.\",\n      \"method\": \"siRNA knockdown, Western blot, full-length cDNA cloning, sequence analysis, endothelial tube formation assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — identifies the long isoform by cloning and links it to tube formation via KD, but functional mechanism is not fully resolved; negative finding on CRAC channel role is also established\",\n      \"pmids\": [\"25475730\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CRACR2A promotes Th1 differentiation and effector function of Th17 cells downstream of TCR signaling: conditional T cell-specific deletion of CRACR2A in mice reduced Th1 responses to LCMV infection and conferred resistance to EAE, with impaired Ca2+/NFAT and JNK/AP1 pathway activation due to deficient TCR signal transmission.\",\n      \"method\": \"Conditional knockout mice (T cell-specific Cre), viral infection model, EAE model, transcript analysis, Ca2+ and JNK signaling assays\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean conditional KO with multiple in vivo disease models and defined signaling readouts, corroborated by prior mechanistic work from same group\",\n      \"pmids\": [\"29987160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Biallelic loss-of-function mutations in CRACR2A cause late-onset combined immunodeficiency in humans. Patient T cells show reduced CRACR2A protein, dampened SOCE (Ca2+ entry), and reduced JNK phosphorylation. Reconstitution experiments in CRACR2A-deleted T cells showed that the E278D allele specifically impairs SOCE/cytokine production (not JNK), while the R144G/E300* truncation impairs both JNK phosphorylation and SOCE, establishing allele-specific separation of CRACR2A's two signaling functions.\",\n      \"method\": \"Patient allele characterization, CRACR2A KO T cell reconstitution with individual mutant alleles, SOCE measurement (Ca2+ imaging), JNK phosphorylation assay, cytokine production assay\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — allele-specific reconstitution in KO T cells with multiple orthogonal readouts (SOCE, JNK, cytokines) rigorously separates two molecular functions\",\n      \"pmids\": [\"34908525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Affinity purification coupled to LC-MS/MS identified dynein and Na+/K+ ATPase subunit alpha 1 (ATP1α1) as validated binding partners (effectors) of GTP-bound Rab46 (CRACR2A) in endothelial cells, confirmed by biochemical and imaging approaches.\",\n      \"method\": \"Affinity purification with constitutively active vs. inactive GFP-Rab46, LC-MS/MS, biochemical validation, imaging\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — affinity purification with nucleotide-locked mutants plus biochemical validation, but limited mechanistic follow-up on ATP1α1 interaction\",\n      \"pmids\": [\"33603063\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In neutrophils, CRACR2A rapidly associates with STIM1 after agonist stimulation and facilitates Ca2+ mobilization, which increases the ligand-binding function of β2 integrin and promotes neutrophil adhesion, crawling, and transmigration. A palmitoylated 20-mer peptide from the coiled-coil region of CRACR2A blocks STIM1-CRACR2A interaction, inhibiting Ca2+ mobilization and β2 integrin activation.\",\n      \"method\": \"Myeloid-specific Cracr2a conditional KO mice, CRISPR/Cas9 KO in dHL-60 cells, CRACR2A rescue overexpression, immunoprecipitation of CRACR2A-STIM1 complex, cytosolic Ca2+ mobilization assay, flow cytometry for integrin activation, intravital microscopy, blocking peptide experiments\",\n      \"journal\": \"Circulation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO mice plus CRISPR KO plus rescue plus peptide inhibitor plus multiple functional readouts (Co-IP, Ca2+, integrin activation, intravital microscopy)\",\n      \"pmids\": [\"39601147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Ca2+ released specifically from TPC2 (two-pore channel 2) endolysosomal channels is required for the EF-hand-dependent detachment of Rab46 (CRACR2A) from the MTOC, thereby enabling secretion of Ang2-positive Weibel-Palade bodies in endothelial cells. Pharmacological TPC2 inhibition increased Rab46 clustering at the MTOC and decreased Ang2 secretion, while a TPC2 agonist had the opposite effect.\",\n      \"method\": \"Ca2+ imaging, high-resolution light microscopy, pharmacological inhibition/activation of TPC2 (Ned19, tetrandrine, TPC2-A1-N), Rab46 localization at MTOC, Ang2 secretion assay\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological approach with imaging and secretion readouts; preprint, single lab, no genetic confirmation of TPC2 specificity\",\n      \"pmids\": [\"bio_10.1101_2025.03.03.641167\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"CRACR2A (EFCAB4B) is a multifunctional Ca2+-sensing protein expressed as a short isoform (EF-hand CRAC channel regulator) and a long isoform (Rab46, a large Rab GTPase) that: (1) forms a Ca2+-dissociable ternary complex with Orai1 and STIM1 to stabilize CRAC channel clusters and promote store-operated Ca2+ entry; (2) acts as a lymphocyte-specific vesicle-associated large Rab GTPase whose PRD-Vav1 interaction drives vesicle translocation to the immunological synapse to activate Ca2+/NFAT and JNK/AP1 TCR signaling; (3) functions as a calcium-activated dynein adaptor, coupling EF-hand Ca2+ sensing to dynein-mediated retrograde endocytic transport; (4) in endothelial cells, integrates G protein and Ca2+ signals (including TPC2-derived Ca2+) to control selective Weibel-Palade body trafficking via dynein; and (5) in neutrophils, associates with STIM1 after agonist stimulation to facilitate Ca2+ mobilization and β2 integrin activation, promoting neutrophil recruitment.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CRACR2A (EFCAB4B) is a Ca2+-sensing protein, expressed as a short EF-hand isoform and a long large-Rab-GTPase isoform (Rab46), that couples calcium signaling to two distinct effector outputs: store-operated Ca2+ entry and dynein-based vesicle transport [#0, #2, #3]. The short isoform directly binds both Orai1 and STIM1 to form a Ca2+-dissociable ternary complex that stabilizes Orai1/STIM1 co-clustering and supports CRAC channel function; constitutive (Ca2+-insensitive) EF-hand mutants drive aberrant STIM1 clustering, Ca2+ overload, and cell death [#0]. In lymphocytes, a large isoform bearing EF-hand motifs, a proline-rich domain (PRD), and a prenylated Rab GTPase domain localizes to Golgi-proximal vesicles whose PRD–Vav1 interaction drives translocation to the immunological synapse, transmitting TCR signals through the Ca2+/NFAT and JNK/AP1 pathways to support Th1 and Th17 effector responses [#1, #5]. Mechanistically, CRACR2A functions as a calcium-activated dynein adaptor: its coiled-coil domain assembles dynein and dynactin into a motile complex and its EF-hand domain confers Ca2+-dependent activation of motility, linking Ca2+ rises to retrograde endocytic transport [#2]. In endothelial cells the Rab46 isoform integrates histamine-driven G protein and Ca2+ signals to selectively route a P-selectin-negative Weibel-Palade body subset to the MTOC via dynein, with EF-hand Ca2+ binding triggering subsequent dispersal and selective cargo release [#3, #9]. In neutrophils CRACR2A associates with STIM1 after agonist stimulation to drive Ca2+ mobilization and β2 integrin activation, promoting recruitment [#8]. Biallelic loss-of-function mutations in CRACR2A cause late-onset combined immunodeficiency, and allele-specific reconstitution separates its SOCE function from its JNK-activating function [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established CRACR2A as a Ca2+-sensing regulator of CRAC channels by showing it bridges Orai1 and STIM1 in a complex that disassembles when Ca2+ rises.\",\n      \"evidence\": \"Affinity purification, reciprocal Co-IP, siRNA knockdown, and EF-hand mutagenesis with confocal cluster imaging\",\n      \"pmids\": [\"20418871\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the ternary complex not resolved\", \"Did not address the existence of a second large isoform\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed that the CRACR2A locus produces a long endothelial isoform (Rab46) with an additional Rab GTPase domain that is functionally distinct from the CRAC-channel-regulating short form.\",\n      \"evidence\": \"cDNA cloning, Western blot, siRNA knockdown, and endothelial tube formation assay\",\n      \"pmids\": [\"25475730\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of contribution to tube formation unresolved\", \"GTPase domain function not yet characterized\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Defined the lymphocyte large isoform as a vesicle-associated Rab GTPase that uses a PRD-Vav1 interaction to deliver TCR signaling to the immunological synapse, positioning CRACR2A in NFAT and JNK pathways.\",\n      \"evidence\": \"Knockout mice, GTP-binding/prenylation mutants, CRACR2A-PRD/Vav1 Co-IP, live-cell vesicle imaging, and signaling assays\",\n      \"pmids\": [\"27016526\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of vesicle cargo at the synapse not defined\", \"How Vav1 GEF activity feeds back on CRACR2A unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated the physiological consequence of CRACR2A loss in T cells, linking it to Th1/Th17 effector function and in vivo immune responses.\",\n      \"evidence\": \"T cell-specific conditional knockout mice in LCMV and EAE models with Ca2+/NFAT and JNK readouts\",\n      \"pmids\": [\"29987160\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not separate the SOCE versus JNK contributions to disease phenotypes\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Established the core molecular mechanism: CRACR2A is a calcium-activated dynein adaptor whose coiled-coil assembles dynein-dynactin and whose EF-hand confers Ca2+-dependent motility, mediating endocytic transport.\",\n      \"evidence\": \"In vitro dynein motility reconstitution plus Ca2+ manipulation and live-cell puncta imaging in Jurkat T cells\",\n      \"pmids\": [\"30814157\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cargo of the endocytic transport process not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Showed that the Rab46 isoform performs selective organelle sorting in endothelium, using dynein for MTOC retrograde transport and EF-hand Ca2+ sensing to disperse a specific WPB subset.\",\n      \"evidence\": \"Nucleotide-locked Rab46 mutants, dynein inhibition, Ca2+ imaging, and selective cargo release assays\",\n      \"pmids\": [\"31092558\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Source of the dispersal-triggering Ca2+ not yet defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified the GTP-bound Rab46 effector proteome, confirming dynein and adding Na+/K+ ATPase subunit ATP1α1 as direct partners.\",\n      \"evidence\": \"Affinity purification of constitutively active vs inactive GFP-Rab46 with LC-MS/MS and biochemical/imaging validation\",\n      \"pmids\": [\"33603063\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional role of the ATP1α1 interaction not pursued\", \"No mechanistic follow-up\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Proved CRACR2A is disease-relevant in humans and genetically dissected its two signaling functions via allele-specific reconstitution.\",\n      \"evidence\": \"Patient allele characterization and CRACR2A-KO T cell reconstitution with SOCE, JNK, and cytokine readouts\",\n      \"pmids\": [\"34908525\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis for the E278D SOCE-specific defect not defined\", \"Spectrum of clinical phenotypes incompletely mapped\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended CRACR2A function to innate immunity, showing a STIM1-coupled Ca2+ mobilization role that activates β2 integrin and drives neutrophil recruitment, and provided a peptide inhibitor.\",\n      \"evidence\": \"Myeloid-specific KO mice, CRISPR KO in dHL-60 cells, rescue, CRACR2A-STIM1 Co-IP, Ca2+ and integrin assays, intravital microscopy, and a coiled-coil blocking peptide\",\n      \"pmids\": [\"39601147\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether neutrophil function uses the short or large isoform not fully resolved\", \"Relationship to dynein-adaptor activity in neutrophils unaddressed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Pinpointed TPC2 endolysosomal Ca2+ as the specific signal driving EF-hand-dependent Rab46 detachment from the MTOC to enable Ang2-positive WPB secretion.\",\n      \"evidence\": \"Ca2+ imaging, high-resolution microscopy, pharmacological TPC2 inhibition/activation, and Ang2 secretion assays (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.03.03.641167\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No genetic confirmation of TPC2 specificity\", \"Single lab, preprint\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the short CRAC-regulatory isoform and the large dynein-adaptor/Rab isoform are differentially deployed across cell types, and whether their functions intersect within a single cell, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified structural model linking EF-hand Ca2+ sensing, coiled-coil dynein assembly, and Rab GTPase activity\", \"Isoform-specific upstream regulation not mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140299\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [1, 3, 7]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3, 9]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 5, 6, 8]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3, 9]}\n    ],\n    \"complexes\": [\n      \"Orai1-STIM1-CRACR2A ternary CRAC complex\",\n      \"dynein-dynactin motile complex\"\n    ],\n    \"partners\": [\n      \"ORAI1\",\n      \"STIM1\",\n      \"VAV1\",\n      \"ATP1A1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":7,"faith_pct":85.71428571428571}}