| 2003 |
Nectin-2 (CD112) was identified as a specific cell surface ligand for the NK activating receptor DNAM-1 (CD226). Protein purification, tryptic digestion, and mass spectrometry identified Nectin-2 as the 65/60 kDa molecule recognized by mAbs that blocked NK cytotoxicity. PVR-Fc and Nectin-2-Fc soluble hybrid molecules directly stained DNAM-1-transfected COS-7 cells, and Nectin-2 expression in cell transfectants enhanced NK-mediated lysis in a DNAM-1-dependent manner. |
Protein purification, mass spectrometry, soluble Fc-fusion binding assays, cell transfection, NK cytotoxicity assays with mAb blocking |
The Journal of experimental medicine |
High |
12913096
|
| 1998 |
Nectin-2 (HveB/Prr2) functions as a herpesvirus entry mediator, conferring susceptibility to infection by HSV-1 mutants unable to use HVEM, HSV-2, and pseudorabies virus, but not wild-type HSV-1. Anti-HveB antibodies blocked infection of HveB-expressing cells. |
cDNA expression library screen in CHO cells, antibody blocking of infection, cell transfection-based entry assay |
Virology |
High |
9657005
|
| 2004 |
DNAM-1 (CD226) functionally interacts with both CD155 (PVR) and CD112 (nectin-2) as ligands. Ectopic expression of CD112 rendered mouse T cells more susceptible to IL-2-activated T and NK cell-mediated cytotoxicity, which was inhibited by anti-CD226 mAb. Although binding affinities of soluble CD226 to CD155 and CD112 were comparable, homophilic interaction of cell-surface CD112 adversely affects CD226 binding to CD112. |
Cell transfection, NK/T cell cytotoxicity assays, mAb blocking, soluble receptor binding affinity measurements |
International immunology |
High |
15039383
|
| 2000 |
Disruption of both alleles of murine nectin-2 gene resulted in sterile males with morphologically aberrant spermatozoa showing defects in nuclear and cytoskeletal morphology and mitochondrial localization. Nectin-2 interacts with l-afadin (an F-actin-binding protein) as a component of cell-cell adherens junctions and is expressed in testes only during later stages of spermatogenesis, indicating a role in late-stage germ cell development and cytoskeletal organization during spermiogenesis. |
Knockout mouse generation, morphological analysis, immunolocalization, genetic epistasis via null phenotype |
Molecular and cellular biology |
High |
10733589
|
| 2000 |
Nectin-2 alpha and delta splice variants (differing in transmembrane/cytoplasmic regions but sharing ectodomain) both mediate HSV-1 mutant (carrying L25P substitution in gD) and HSV-2 entry at ~100-fold lower efficiency than HveC/HIgR. gD from HSV-1(U21) bound in vitro soluble forms of nectin-2, with weaker association than to HveC/HIgR. The major functional region for virus entry mapped to the V domain at the N-terminus. |
Cell transfection entry assay, in vitro gD binding assay, domain mapping |
Journal of virology |
High |
10627537
|
| 2000 |
Nectin-2 alpha mediates direct cell-to-cell spread of HSV-1 mutant virus (carrying L25P gD substitution) but not wild-type HSV-1, consistent with its entry receptor specificity. An antibody to nectin-1 blocked cell-to-cell spread, and wild-type virus did not spread from receptor-positive to receptor-negative cells, demonstrating that Ig-like receptors (nectin-1 and nectin-2) are required for both virus entry and cell-to-cell spread. |
Plaque formation assay in receptor-expressing J cells, mAb blocking of spread, genetic analysis of receptor requirements |
Journal of virology |
High |
10729168
|
| 2003 |
Loss of nectin-2 at Sertoli-spermatid junctions causes male infertility. Nectin-2-deficient spermatozoa show severe head and midpiece malformations, 4-fold reduced migration to oviducts, 6-fold reduced binding to zona-intact oocytes, and failure to penetrate zona-free hamster oocytes. Sertoli-spermatid junctions in nectin-2-null mice lack the actin-bundling protein espin, indicating ectoplasmic specializations fail to form. Nectin-2 is produced exclusively by Sertoli cells and forms a heterotypic adhesion complex with nectin-3 on elongated spermatids. |
Knockout mouse (LacZ knockin), scanning electron microscopy, in vitro fertilization assays, LacZ reporter expression analysis, immunolocalization |
Biology of reproduction |
High |
12801998
|
| 2001 |
Specific amino acid sequences in the V domain of human nectin-2 are critical for HSV entry activity. Chimeric molecules between human and mouse nectin-2 showed that replacing amino acids 75-81 or residue 89 of mouse nectin-2 with human residues transferred HSV-1/Rid and HSV-2 entry activity. Replacement of residue 89 of human nectin-2 (M89F) eliminated HSV entry activity. This region is homologous to the HIV-binding region of CD4 and the poliovirus-binding region of CD155. |
Chimeric receptor construction, cell transfection entry assay, mutagenesis |
Journal of virology |
High |
11602758
|
| 2003 |
N-terminal sequences of HSV gD govern functional interactions with nectin-2, HVEM, and 3-O-sulfated heparan sulfate, but are not required for nectin-1 interactions. Deletions overlapping HVEM contact regions (aa 7-15 and 24-32) severely reduced cell fusion and binding with all receptors except nectin-1. L25P, Q27P, and Q27R substitutions in HSV-1 gD enhanced fusion with human nectin-2 (ordinarily low for wt HSV-1 gD) while the same substitutions in HSV-2 gD were without effect on the already high fusion level. |
Cell fusion assay, soluble gD:Fc binding to receptor-expressing cells, mutagenesis (substitutions and deletions in gD N-terminus) |
Journal of virology |
High |
12915538
|
| 2010 |
Human cytomegalovirus UL141 promotes downregulation of CD112 (nectin-2) via proteasome-mediated degradation by 48 h post-infection, thereby removing both DNAM-1 activating ligands (CD155 and CD112) from the cell surface. UL141 alone is sufficient to retain CD155 in the ER, but requires additional HCMV-encoded functions to suppress CD112 expression. |
HCMV infection with UL141 deletion mutant, proteasome inhibitor treatment, flow cytometry for surface expression, genetic rescue with UL141 deletion |
The Journal of general virology |
High |
20410314
|
| 2012 |
Crystal structure of the nectin-2 V-set domain (nectin-2v) at 1.85 Å resolution reveals a perpendicular homodimer arrangement. Disruption of the homodimeric interface by mutagenesis abolished homodimer formation and simultaneously eliminated DNAM-1 binding, indicating that homodimerization or engagement of the homodimeric interface is required for DNAM-1 interaction. |
X-ray crystallography (1.85 Å), mutagenesis, surface plasmon resonance, tetramer cell staining, biochemical characterization |
Journal of immunology |
High |
22547693
|
| 2012 |
Crystal structure of the nectin-2 homodimer at 1.3 Å resolution reveals the structural basis for homophilic and heterophilic interactions. Mutagenesis of charged residues at the dimer interface identifies them as major determinants of binding affinities, and provides mechanistic explanation for stronger heterophilic versus weaker homophilic interactions among nectin family members. |
X-ray crystallography (1.3 Å), complementary mutagenesis, biochemical binding assays |
Proceedings of the National Academy of Sciences |
High |
22927415
|
| 2010 |
Nectin-2 physically interacts with N-cadherin through their extracellular domains, and they cooperatively enhance apical constriction by driving F-actin accumulation at apical cell surfaces during Xenopus neural tube morphogenesis. The intracellular afadin-binding motif of nectin-2 was not required for ectopic apical constriction induction. Nectin-2 knockdown impaired neural fold formation by attenuating F-actin accumulation, while overexpression in non-neural ectoderm induced ectopic apical constrictions. |
Xenopus morpholino knockdown, overexpression of domain-deleted nectin-2, co-immunoprecipitation of extracellular domains, live imaging of F-actin |
Development |
High |
20332149
|
| 2017 |
TIGIT binds to the membrane-distal V-set immunoglobulin domain of nectin-2 with an affinity of 6 μM. Crystal structure of TIGIT bound to the first Ig domain of nectin-2 showed that TIGIT and nectin-2 dock using the same molecular surface (lock-and-key binding motif) used in nectin/nectin homotypic interactions. TIGIT/nectin-2 binding disrupts pre-assembled nectin-2 oligomers. Mutagenesis identified an 'aromatic key' of nectin-2 as critical for TIGIT interaction, while the C-C' loop of nectin-2 dictates the TIGIT binding hierarchy. |
X-ray crystallography of TIGIT:nectin-2 complex, SPR binding affinity measurements, mutagenesis, disruption of nectin-2 oligomers assay |
The Journal of biological chemistry |
High |
28515320
|
| 2016 |
TIGIT engages nectin-2 using a canonical immunoglobulin-like dimer interface. Biophysical studies showed TIGIT is monomeric in solution but dimerizes at high concentrations. Biochemical mutagenesis mapped the nectin-2 binding interface on TIGIT, defining the structural and biochemical determinants for TIGIT:nectin-2 recognition. |
X-ray crystallography of TIGIT ectodomain, biophysical characterization (SPR), mutagenesis, model of TIGIT:nectin-2 complex |
Molecular immunology |
High |
27978489
|
| 2006 |
DNAM-1 (CD226) and its ligand Nectin-2 (CD112) are expressed on human mast cells and eosinophils. Engagement of CD226 on mast cells augments FcεRI-dependent degranulation through a pathway involving Fyn, LAT, PLCγ2, and CD18. This costimulatory pathway is completely inhibited by IRp60 (CD300a) inhibitory receptor. Blocking CD112 on eosinophils normalized the hyperactivation of mast cells co-cultured with eosinophils. |
Flow cytometry, mAb stimulation/blocking, degranulation assay, inhibitor analysis of signaling pathway, co-culture with blocking antibodies |
The Journal of biological chemistry |
Medium |
16831868
|
| 2009 |
Nectin-2 deficiency in mice leads to cardiac fibrosis and dysfunction under chronic pressure overload (ascending aorta banding) but not under physiological conditions. In banded nectin-2 knockout mice, intercalated discs were disrupted, myofibrils disorganized, and cardiac myocyte apoptosis increased. Akt phosphorylation remained lower, while JNK and p38 MAPK were hyperphosphorylated compared to wild-type, revealing nectin-2-dependent signaling for cardiac protection. |
Knockout mouse, ascending aorta banding model, echocardiography, histology, immunoblotting for signaling kinases, TUNEL assay |
Hypertension |
High |
19667252
|
| 2013 |
Nectin-3 (CD113) expressed on T lymphocytes trans-interacts with Nectin-2 (CD112) on endothelial cells at high endothelial venules to promote lymphocyte transendothelial migration. A soluble form of Nectin-3 binds to Nectin-2 at EC junctions, and blocking Nectin-2 trans-interactions with mAbs abolished soluble Nectin-3 binding. Blocking either Nectin-3 on lymphocytes or Nectin-2 on ECs inhibited lymphocyte extravasation in vitro. |
Soluble receptor binding assay, mAb blocking of trans-interactions, in vitro transendothelial migration assay |
PloS one |
High |
24116228
|
| 2014 |
The alphaherpesvirus gD glycoprotein (from PRV and HSV-2) induces degradation and downregulation of CD112 (nectin-2) from the cell surface during infection or transfection, reducing DNAM-1 binding to infected/transfected cells and suppressing NK cell degranulation and lysis. This identifies a previously uncharacterized alphaherpesvirus NK evasion strategy. |
Virus infection/transfection, flow cytometry for surface CD112, DNAM-1-Fc binding assay, NK cell degranulation and cytotoxicity assays |
Proceedings of the National Academy of Sciences |
High |
25352670
|
| 2016 |
Nectin-2 (CD112) regulates endothelial cell biology in vivo: CD112-deficient mice show enhanced blood vessel coverage in retina and spleen. In vitro, CD112 blockade enhanced endothelial tube formation and cell migration. CD112 knockdown enhanced tube formation, cell migration, and proliferation with p-Erk activation, and led to compensatory upregulation of Nectin-3 and Necl-4 which promote VEGFR signaling. CD112 also regulates T cell entry into the spleen in vivo. |
CD112-knockout mice, retinal and spleen vasculature analysis, in vitro tube formation and migration assays, siRNA knockdown, flow cytometry, signaling analysis |
PloS one |
Medium |
27676263
|
| 2019 |
Ubiquitination of Nectin2 promotes its intracellular retention and proteasomal degradation, limiting surface expression. Inhibition of the ubiquitin-proteasome pathway results in increased Nectin2 surface expression and enhances tumor cell susceptibility to NK cell cytotoxicity. Nectin2 is predominantly expressed in cytoplasmic pools on tumor cell lines. |
Ubiquitin pathway inhibitors, flow cytometry for surface vs. total expression, NK cell cytotoxicity assays, subcellular fractionation |
European journal of immunology |
Medium |
30888046
|
| 2016 |
Nectin-2δ splice variant is selectively expressed in astrocytes and localizes to perivascular astrocytic endfoot processes facing blood vessel basement membranes, while nectin-2α is expressed in both neurons and astrocytes. Genetic ablation of nectin-2 caused degeneration of astrocytic perivascular endfoot processes and neurons in the cerebral cortex, revealing a critical role for nectin-2 in brain homeostasis. |
Nectin-2 knockout mouse, immunofluorescence, confocal microscopy, cell-type specific fractionation, electron microscopy of endfoot processes |
Brain research |
High |
27545667
|
| 2006 |
Nectin-2 gene transcription in testicular cells is controlled by cooperative interaction of Sp1, CREB, and AP-1 (c-Jun) transcription factors. The minimal mouse nectin-2 promoter (nucleotides -316 to -211) contains two Sp1 motifs and one CRE motif that synergize to regulate transcription. In vivo, CREB, c-Jun, and Sp1 family proteins are bound to the nectin-2 promoter. Cyclic expression of CREB coincides with adherens junction restructuring between Sertoli cells and germ cells. |
Transient transfection/promoter reporter assay, EMSA, chromatin immunoprecipitation (ChIP), siRNA knockdown, overexpression |
Journal of cellular physiology |
High |
16250013
|
| 2014 |
Cadmium suppresses nectin-2 expression at both transcriptional and post-translational levels in testicular cells. Cadmium induces nectin-2 protein degradation via clathrin-dependent endocytosis, promotes nectin-2 internalization, and directly represses nectin-2 transcription by inhibiting binding of positive transcriptional regulators (Sp1, CREB) to the nectin-2 promoter, contributing to blood-testis barrier disruption. |
Endocytosis inhibitors, shRNA knockdown, siRNA, immunofluorescence, EMSA, ChIP, flow cytometry-based endocytosis assay |
Biochimica et biophysica acta |
High |
25046863
|
| 2018 |
Nectin-2 directly interacts with the extracellular domain of N-cadherin with a KD of 3.5 ± 0.6 μM, as demonstrated by surface plasmon resonance. Structural analysis of homodimeric structures and molecular docking identified the binding interface. This provides a direct, adaptor-independent mechanism by which nectins can recruit cadherins to sites of adherens junction. |
Surface plasmon resonance (SPR), structural analysis of existing crystal structures, molecular docking, complementary mutagenesis |
Proteins |
Medium |
30183103
|
| 2022 |
Nectin-2 acts as a viral entry mediator for Human Herpesvirus 6B (HHV-6B). Nectin-2-transduced T cells became permissive for HHV-6B infection, and nectin-2 knockout in parotid-derived HSY cells significantly reduced HHV-6B entry. HHV-6B glycoprotein B (gB) interacted specifically with the nectin-2 V-set domain. |
Nectin-2 gene transduction into T cells, nectin-2 knockout via CRISPR, virus infection assays, pulldown/binding assay between gB and nectin-2 V-set domain |
Viruses |
High |
35062364
|
| 2024 |
Crystal structure of the PVRIG (CD112R):Nectin-2 complex at undisclosed resolution identified a unique CC' loop in PVRIG that complements a double-lock-and-key binding mode and contributes to its high affinity for Nectin-2. Charged residues in the F-strands explain PVRIG selectivity for Nectin-2 over Necl-5. CD112R was found to disrupt CD112 homodimerization in an antiparallel binding mode. |
X-ray crystallography of PVRIG:Nectin-2 complex, comparative binding affinity measurements, structural analysis |
Structure |
High |
38626767
|
| 2025 |
Crystal structure of the CD112:CD112R (Nectin-2:PVRIG) complex at 2.2 Å resolution reveals an antiparallel, lock-and-key binding mode in which CD112R disrupts CD112 homodimerization. Structure-guided directed evolution produced CD112R mutants with increased CD112 binding affinity; the highest-affinity variant (CD112RIVE) potently inhibits CD112-CD112R interactions as a soluble trap. |
X-ray crystallography (2.2 Å), directed evolution, SPR binding affinity measurement, functional T cell activation and cytotoxicity assays with CAR-T and T cell engager constructs |
Molecular therapy |
High |
40285356
|
| 2002 |
Mutations in three equivalent regions of the N-terminal V-like domain of nectin-2 revealed distinct structural requirements for HSV entry, PRV/BHV-1 entry, and homotypic/heterotypic nectin-nectin trans-interactions. Region I mutations (impairing HSV entry) did not reduce homotypic trans-interactions for nectin-2. Region III mutations that reduced homotypic trans-interactions of nectin-2 also impaired PRV and BHV-1 entry activity. Effects on gD binding did not necessarily correlate with functional entry. |
Mutagenesis of nectin-2 V domain, cell transfection entry assay with multiple alphaherpesvirus strains, binding assay to gD |
Journal of virology |
High |
12438620
|
| 2025 |
ST6GalNAc-I sialylates NECTIN2 in lung adenocarcinoma cells, as shown by proteomic and biochemical analysis. This sialylation promotes T cell dysfunction via the NECTIN2-TIGIT axis. ST6GalNAc-I-deficient tumor cells co-cultured with T cells were more susceptible to T cell-mediated killing, and mice injected with St6galnac-I-knockdown cells showed reduced Nectin2/Tigit-associated immunosuppression. |
Proteomics, biochemical co-localization, syngeneic mouse model, KD/KO co-culture cytotoxicity assay |
The Journal of clinical investigation |
Medium |
40371640
|
| 2006 |
Nectin-2-mediated HSV-1 ANG path (syncytial strain) entry into CHO cells occurs via pH-independent fusion at the plasma membrane, whereas nectin-1 directs the same strain to a pH-dependent endocytic pathway. HSV-1 ANG path virions induced rapid fusion-from-without (FFWO) exclusively in CHO-nectin-2 cells, not in CHO-nectin-1 or receptor-negative cells, demonstrating that receptor identity (nectin-2 vs. nectin-1) determines the entry pathway. |
Endocytosis/acidification inhibitors, kinetics of entry measurement, FFWO assay, CHO cell transfectants expressing distinct receptors |
Virology journal |
High |
17192179
|
| 1999 |
Murine nectin-2 (mHveB), the mouse homolog of human HveB, mediates entry of porcine pseudorabies virus (PRV) but not HSV-1 or HSV-2. Anti-mHveB antibody and soluble mHveB ectodomain inhibited mHveB-dependent PRV entry. Co-expression of mHveB with PRV gD (but not HSV-1 gD) caused interference with entry, suggesting direct interaction of PRV gD with mHveB during viral entry. |
Cell transfection entry assay, antibody blocking, interference assay with viral gD co-expression, RT-PCR |
Journal of virology |
High |
10196354
|
| 2021 |
VSIG2 specifically binds to Nectin-2 (and not to PD-1 or CTLA-4) and functions as an immunosuppressive ligand present on antigen-presenting cells. VSIG2-Nectin-2 binding strongly inhibits T cell activation and proliferation. This interaction regulates the STAT1/IRF1/GBP2 signaling pathway in T cells. |
Direct binding assay (VSIG2 vs. known immune receptors), co-culture T cell activation/proliferation assays, signaling pathway analysis, in vivo mouse EAE model, anti-tumor assay |
Journal of neuroinflammation |
Medium |
41350674
|