Affinage

NCK2

Cytoplasmic protein NCK2 · UniProt O43639

Length
380 aa
Mass
42.9 kDa
Annotated
2026-06-10
42 papers in source corpus 28 papers cited in narrative 27 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NCK2 is a modular SH2/SH3 adaptor protein that couples activated receptor tyrosine kinases and adhesion receptors to actin-regulatory effectors, thereby controlling cytoskeletal dynamics, cell migration, neuronal morphogenesis, and cytokinesis (PMID:9843575, PMID:11027258, PMID:19242519). Built from three N-terminal SH3 domains and a single C-terminal SH2 domain, it engages receptors through phosphotyrosine-dependent SH2 binding — EGFR, PDGFR-β at Tyr-1009, TrkB, and the integrin scaffold FAK at phospho-Tyr397 — while its tandem SH3 domains recruit downstream effectors including DOCK180, PAK3, IRS-1, paxillin, GIT1, and Robo (PMID:9843575, PMID:11027258, PMID:11240126, PMID:11950595, PMID:12074588, PMID:16636066, PMID:16752908, PMID:21600986). Its three SH3 modules carry distinct binding preferences (SH3-2 favoring APxxPxR and SH3-3 favoring PxAPxR motifs, with tandem domains binding cooperatively), and although the NCK2 SH2 domain is structurally distinguished by a unique C-terminal antiparallel β-sheet, its phosphopeptide specificity is essentially indistinguishable from NCK1, implying that functional divergence between the paralogs arises largely from SH3-domain selectivity and differential regulation (PMID:11240126, PMID:15764601, PMID:16636066, PMID:16752908). Through these interactions NCK2 directs specific small-GTPase outputs: it transmits PDGFR-β Tyr-1009 signals to RhoA-driven stress fibers (whereas NCK1 routes Tyr-751 to Cdc42) and is recruited by the integrin adaptor Cas to activate Cdc42 for polarized migration (PMID:19242519, PMID:20637038). NCK2 has non-redundant, NCK1-independent roles in NGF- and Slit1-induced neurite outgrowth — in part by binding paxillin and protecting it from proteasomal degradation — in PAK3-dependent down-regulation of synaptic transmission, in Eph-mediated motor axon guidance via α2-chimaerin, and in completing cytokinetic abscission, where its SH2 domain is required for proper midbody localization of AURKB, PLK1, and ECT2 (PMID:17591694, PMID:21949127, PMID:21600986, PMID:30002203, PMID:30016580). NCK2 abundance is set post-translationally by CBL-mediated ubiquitin-proteasomal degradation and transcriptionally by KLF4, and unlike NCK1 it lacks the Lys178 c-Cbl ubiquitination acceptor site, providing a molecular basis for paralog-specific stability (PMID:24287595, PMID:36504051, PMID:38583810).

Mechanistic history

Synthesis pass · year-by-year structured walk · 21 steps
  1. 1998 High

    Established NCK2 as a distinct SH2/SH3 adaptor and defined its first domain-specific receptor and scaffold interactions, framing it as a modular signaling linker rather than a redundant NCK1 copy.

    Evidence Yeast two-hybrid, Co-IP, GST pulldown and domain mutagenesis mapping PINCH, EGFR, PDGFR-β and IRS-1 contacts

    PMID:9843575

    Open questions at the time
    • Did not establish functional output of these interactions
    • Cytoskeletal association observed but mechanism undefined
  2. 2000 High

    Demonstrated NCK2 has a function not shared by NCK1 in PDGF-induced actin remodeling, providing the first evidence of paralog-specific signaling.

    Evidence Isoform-specific antibody microinjection, overexpression, and SH2/SH3 mutants in fibroblasts, mapping NCK2 to PDGFR-β Tyr-1009

    PMID:11027258

    Open questions at the time
    • Effector linking NCK2 to actin not yet identified
    • Relationship to Rac1 inferred from epistasis only
  3. 2001 High

    Resolved how NCK2 SH3 domains achieve high-affinity, cooperative effector binding (DOCK180, IRS-1), explaining how tandem domains generate signaling specificity and avidity.

    Evidence SPR kinetics, GST pulldown, and point mutagenesis of tandem SH3 domains and partner motifs

    PMID:11171109 PMID:11240126

    Open questions at the time
    • Cellular consequence of DOCK180 recruitment not tested here
    • In vivo stoichiometry of cooperative binding unknown
  4. 2002 Medium

    Connected NCK2 to focal-adhesion (FAK) and neurotrophin (TrkB) signaling, broadening its receptor repertoire to adhesion and neuronal contexts.

    Evidence Co-IP, phospho-Tyr397-dependent SH2 binding, co-localization and motility assays; BDNF-dependent TrkB Co-IP from cortical neurons

    PMID:11950595 PMID:12074588

    Open questions at the time
    • Opposing motility effects of full-length vs SH2-only NCK2 not mechanistically resolved
    • TrkB downstream pathway not defined
  5. 2005 High

    Provided the structural basis for NCK2 SH2 recognition and revealed a unique fold feature plus multimodal ephrinB2 phosphopeptide binding.

    Evidence NMR solution structure and HSQC titrations with phospho-ephrinB2 fragments

    PMID:15764601

    Open questions at the time
    • Functional role of the unique C-terminal β-sheet untested
    • ephrinB2-NCK2 cellular signaling not addressed
  6. 2006 High

    Defined SH3 motif preferences and showed that NCK1 and NCK2 SH2 domains have indistinguishable phosphopeptide specificity, localizing paralog divergence to SH3 domains.

    Evidence X-ray crystallography, ITC, and NMR with Tir, GIT1 and Nogo-A peptides

    PMID:16636066 PMID:16752908

    Open questions at the time
    • Does not explain how indistinguishable SH2 domains produce divergent receptor coupling in cells
    • GIT1/Nogo-A functional roles not tested
  7. 2007 High

    Identified an NCK2-specific mechanism in neurons: stabilizing paxillin against proteasomal degradation to enable neurite outgrowth, establishing a non-adaptor (protein-stability) role.

    Evidence Isoform-specific shRNA, Co-IP, proteasome inhibition, and non-degradable NCK2/paxillin rescue in PC12 and cortical neurons

    PMID:17591694

    Open questions at the time
    • Molecular basis of NCK2-dependent paxillin protection unclear
    • Why effect is neuron-specific not explained
  8. 2009 High

    Showed NCK1 and NCK2 are non-compensating, routing the same receptor to distinct GTPases (NCK2→RhoA/stress fibers at Tyr-1009; NCK1→Cdc42/filopodia at Tyr-751).

    Evidence NCK2-KO MEFs, RNAi, PDGFR-β site mutants, dominant-negative SH3, and GTPase activity assays

    PMID:19242519

    Open questions at the time
    • Intermediate GEF/effector linking NCK2 to RhoA not defined
    • Generality beyond fibroblasts untested
  9. 2010 Medium

    Placed NCK2 downstream of integrin/Cas signaling as a Cdc42 activator for cell polarization, distinguishing it from CrkII-driven Rac1 protrusion.

    Evidence siRNA knockdown, GTPase activity assays, and wound-healing assays

    PMID:20637038

    Open questions at the time
    • Direct NCK2-Cas binding mode not mapped here
    • GEF coupling NCK2 to Cdc42 unknown
  10. 2011 High

    Extended NCK2-specific functions into neuronal guidance, synaptic transmission, and lamellipodial actin assembly through Robo, PAK3, and SKAP2 partners.

    Evidence Isoform-specific knockdown, Co-IP from brain, interfering peptide plus electrophysiology, and leading-edge co-localization

    PMID:20219016 PMID:21600986 PMID:21949127

    Open questions at the time
    • Atypical SH3-mediated Robo binding mode not structurally defined
    • How NCK2-PAK3 down-regulates transmission mechanistically open
  11. 2013 High

    Explained paralog-specific stability: NCK1 is ubiquitinated by c-Cbl at Lys178 while NCK2 lacks the acceptor site, accounting for differential turnover and signaling persistence.

    Evidence Ubiquitination assays, K178R mutagenesis, domain swaps, and RhoA activity assays in podocytes

    PMID:24287595

    Open questions at the time
    • Whether NCK2 has its own degradation pathway not addressed here
    • Functional consequence of NCK2 stability in this context not isolated
  12. 2014 Medium

    Distinguished NCK2 signaling output in T cells, linking it selectively to TCR-induced NFAT activation rather than the Erk/AP-1 axis used by NCK1.

    Evidence Isoform-specific shRNA, luciferase reporters, and phospho-immunoblotting in Jurkat cells

    PMID:24670066

    Open questions at the time
    • Molecular link from NCK2 to NFAT not defined
    • Direct TCR-proximal partners in this pathway not identified
  13. 2016 Medium

    Revealed a metabolic role: NCK2 restrains adipogenesis by suppressing PERK signaling in adipocyte precursors.

    Evidence NCK2-KO mice and in vitro differentiation assays with PERK signaling analysis

    PMID:27325288

    Open questions at the time
    • Direct NCK2-PERK molecular link unestablished
    • Adaptor partners in adipocytes not mapped
  14. 2018 High

    Defined an NCK1-independent requirement for NCK2 in cytokinetic abscission, with its SH2 domain needed for proper midbody localization of AURKB, PLK1 and ECT2, and an NCK2-specific interactome.

    Evidence NCK2-KO MEFs, live imaging, AP-MS and BioID, and SH2-mutant rescue

    PMID:30002203

    Open questions at the time
    • Direct SH2 ligand at the midbody not identified
    • Mechanism positioning AURKB/PLK1/ECT2 via NCK2 unresolved
  15. 2018 Medium

    Showed NCK2 is required for Eph/ephrin-guided motor axon trajectory selection by modulating α2-chimaerin.

    Evidence In ovo electroporation loss/gain-of-function, ephrin neurite assays, and α2-chimaerin epistasis

    PMID:30016580

    Open questions at the time
    • Direct NCK2-α2-chimaerin binding not biochemically mapped
    • Eph receptor coupling specificity unresolved
  16. 2020 High

    Sharpened paralog distinction in endothelium: both adaptors bind PECAM-1 under shear, but only NCK1 (via its first SH3) drives PAK2 activation and permeability, with NCK2 dispensable.

    Evidence Knockout cells, domain swaps, single-point SH3 mutants, in vivo carotid ligation, and permeability/PAK assays

    PMID:32468886

    Open questions at the time
    • NCK2's role at PECAM-1, if any, not defined
    • Why NCK2 SH3 domains fail to recruit PAK here unexplained
  17. 2022 Medium

    Identified CBL as an E3 ligase driving NCK2 ubiquitin-proteasomal degradation, controlling NCK2 abundance and its pro-proliferative/migratory output in cancer.

    Evidence Co-IP, IP-MS, cycloheximide chase, ubiquitination assay, and rescue overexpression in breast cancer cells

    PMID:36504051

    Open questions at the time
    • NCK2 ubiquitination acceptor site not mapped (contrast with NCK1 Lys178)
    • Single-lab finding without in vivo validation
  18. 2023 Medium

    Linked NCK2 to spermatogonial stem cell self-renewal as an OIP5 effector promoting proliferation and survival.

    Evidence Co-IP, IP-MS, GST pulldown, and siRNA functional assays in human SSCs

    PMID:37292517

    Open questions at the time
    • Domain mediating OIP5-NCK2 interaction unmapped
    • Downstream signaling in SSCs undefined
  19. 2024 Medium

    Positioned NCK2 as a scaffold in the EphA4/RhoA axis in olfactory neurons under α-synuclein control, tying its abundance to neuronal projection.

    Evidence α-synuclein-KO mice, immunoprecipitation, iTRAQ-LC-MS, and overexpression rescue

    PMID:39543759

    Open questions at the time
    • Direct NCK2-EphA4/RhoA binding interfaces not mapped
    • How α-synuclein controls NCK2 levels unknown
  20. 2024 Medium

    Identified KLF4 as a direct transcriptional activator of NCK2, embedding NCK2 in a miR-150-3p/KLF4 axis that modulates melanoma drug response.

    Evidence ChIP, promoter reporter assay, and overexpression/knockdown with Western blot

    PMID:38583810

    Open questions at the time
    • Downstream NCK2 effectors in melanoma not identified
    • Single-context regulatory finding
  21. 2025 Medium

    Implicated NCK2 in clathrin-coated pit formation and EGF-driven PI3K/Akt signaling at the plasma membrane, suggesting a role in receptor-proximal endocytic signaling platforms.

    Evidence TIRFM CCP dynamics, Akt phosphorylation, and PI3K localization assays (preprint)

    PMID:bio_10.1101_2025.10.24.684344

    Open questions at the time
    • Preprint, not peer-reviewed
    • NCK2-specific vs shared NCK1 contribution to CCPs not separated
    • Direct PI3K recruitment mechanism unmapped

Open questions

Synthesis pass · forward-looking unresolved questions
  • The molecular events linking NCK2's effector recruitment to specific small-GTPase activation (which GEFs it engages for RhoA vs Cdc42) and the structural/sequence basis for its paralog-specific cellular outputs despite SH2 specificity shared with NCK1 remain unresolved.
  • No GEF directly bridging NCK2 to RhoA/Cdc42 identified
  • Midbody SH2 ligand in cytokinesis unknown
  • NCK2's own ubiquitination acceptor site (vs NCK1 Lys178) unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 4
Localization
GO:0005886 plasma membrane 3 GO:0005856 cytoskeleton 2 GO:0005634 nucleus 1
Pathway
R-HSA-1266738 Developmental Biology 3 R-HSA-162582 Signal Transduction 3 R-HSA-1640170 Cell Cycle 1 R-HSA-5653656 Vesicle-mediated transport 1
Partners
DOCK180FAKIRS1PAK3PAXILLINPDGFRBPINCHROBO

Evidence

Reading pass · 27 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1998 NCK2 (Nck-2) was identified as an adaptor protein comprising three N-terminal SH3 domains and one C-terminal SH2 domain. It interacts with PINCH via the fourth LIM domain of PINCH and the third SH3 domain of NCK2. NCK2 also binds EGF receptor (largely via its SH2 domain, with SH3 domains contributing), PDGF receptor-β (solely via SH2 domain, PDGF-dependent), and IRS-1 (primarily via second and third SH3 domains). A fraction of NCK2 was found associated with the cytoskeleton. Yeast two-hybrid, Co-IP, GST pulldown, domain mutagenesis Molecular biology of the cell High 9843575
2000 NCK2 (Nckbeta) plays a specific role in PDGF-BB-induced actin polymerization. Overexpression of NCK2 but not NCK1 blocks PDGF-stimulated membrane ruffling and lamellipodia. Mutation in either the SH2 or the middle SH3 domain abolishes this effect. NCK2 binds PDGFR-β at Tyr-1009 (distinct from NCK1's Tyr-751 binding site). Anti-NCK2 but not anti-NCK1 microinjection inhibits PDGF-stimulated actin polymerization. Constitutively membrane-bound NCK2 blocks Rac1-L62-induced membrane ruffling, suggesting NCK2 acts in parallel to or downstream of Rac1. Overexpression, dominant-negative mutants, microinjection of isoform-specific antibodies, site-directed mutagenesis Molecular and cellular biology High 11027258
1999 NCK2 (Grb4) associates with receptor tyrosine kinases and SH3-binding proteins PAK, Sos1, and PRK2. NCK2 synergizes with v-Abl and Sos1 to induce Elk-1-dependent gene expression and cooperates with v-Abl to transform NIH 3T3 cells. Co-IP, reporter gene assay, transformation assay The Journal of biological chemistry Medium 10026169
2001 NCK2 interacts with DOCK180 via its second and third SH3 domains. A major binding site maps to DOCK180 residues 1819–1836 (recognized primarily by the third SH3 domain). Two binding events occur with equilibrium dissociation constants of ~415 nM and ~3.24 nM. Both SH3 domains contribute cooperatively, with tandem SH3 domains greatly enhancing binding compared to individual domains alone. Yeast two-hybrid, GST pulldown, surface plasmon resonance, site-directed mutagenesis FEBS letters High 11240126
2001 NCK2 SH3 domains directly interact with IRS-1 in vivo. Multiple SH3 domains (with conserved tryptophan residues critical) enhance complex formation. IRS-1 PTB/SAIN domain and Pre-C-terminal domain (but not PH domain) mediate NCK2 binding. The interaction is direct (occurs in absence of other proteins). Co-IP, GST pulldown, in vitro binding assay, deletion and point mutagenesis The Biochemical journal High 11171109
2002 NCK2 interacts with focal adhesion kinase (FAK) through multiple SH2 and SH3 domains. The SH2-mediated interaction requires phosphorylation of FAK Tyr397. A fraction of NCK2 co-localizes with FAK at the cell periphery in spreading cells. Overexpression of NCK2 modestly decreases cell motility, whereas a SH2-only NCK2 mutant lacking SH3 domains significantly promotes motility. Co-IP, mutagenesis, immunofluorescence co-localization, overexpression motility assay The international journal of biochemistry & cell biology Medium 11950595
2002 BDNF stimulation promotes interaction of NCK2 with the TrkB tyrosine kinase receptor. Tyrosines Y694, Y695, and Y771 in the TrkB intracellular domain are crucial for this interaction. NCK2 was co-precipitated with GST-NCK2 recombinant protein or anti-Nck antibody from BDNF-activated cortical neurons. Yeast two-hybrid, GST pulldown, Co-IP from cortical neurons, mutagenesis Biochemical and biophysical research communications Medium 12074588
2001 NCK2 (Grb4) acts as a nuclear repressor of v-Abl-induced transcriptional activation from AP-1 and SRE promoter elements. This inhibitory activity is independent of direct v-Abl/NCK2 SH2 interaction; a SH2 domain mutant shows even stronger inhibition. The first two SH3 domains primarily mediate inhibitory function. The inhibitory activity is downstream of MEKK1 and JNK. Cell fractionation and fluorescence microscopy revealed that stronger inhibitory SH2 mutants show increased nuclear localization. Reporter gene assay, domain mutagenesis, cell fractionation, fluorescence microscopy The Journal of biological chemistry Medium 11514578
2005 The NMR solution structure of the human NCK2 SH2 domain was determined. It adopts the core SH2 fold but with a unique C-terminal antiparallel β-sheet not previously identified in other SH2 domains. The NCK2 SH2 domain binds three phosphorylated ephrinB2 fragments ([Tyr(P)304], [Tyr(P)316], and [Tyr(P)330]ephrinB2) via different mechanisms and with distinct conformational dynamics. NMR structure determination, NMR titration (HSQC), truncation mutagenesis The Journal of biological chemistry High 15764601
2006 Crystal structures of NCK1 and NCK2 SH2 domains in complex with phosphopeptides from the EPEC protein Tir established that the SH2 domains of NCK1 and NCK2 have essentially indistinguishable phosphopeptide binding specificities. The second SH3 domain of NCK2 prefers APx#PxR motifs and the third SH3 domain prefers PxAPxR. NCK2 SH3 domains bind GIT1 (Arf-GAP) as experimentally confirmed. High-affinity binding of NCK2 SH3-3 to Nogo-A peptide was identified (Kd = 5.7 μM). X-ray crystallography, NMR (HSQC titration), isothermal titration calorimetry, binding assays The Journal of biological chemistry / Biochemistry High 16636066 16752908
2007 NCK2 (Nckbeta), but not NCK1, is required for nerve growth factor-induced neurite outgrowth in PC12 cells and for normal axon/dendrite development in primary cortical neurons. NCK2 knockdown reduces steady-state paxillin levels specifically in neurons (not glia). NCK2 binds strongly to paxillin, preventing its proteasomal degradation. Re-expression of non-degradable NCK2 or forced paxillin expression rescues neuritogenesis in NCK2 knockdown cells. shRNA knockdown, overexpression rescue, Co-IP, proteasome inhibitor treatment, immunoblotting Molecular and cellular biology High 17591694
2009 NCK2 (Nckbeta) and NCK1 (Nckalpha) have non-compensating roles in PDGF-BB-induced dermal fibroblast migration. NCK2 binds PDGFR-β at Tyr-1009 and mediates Rho signaling to induce stress fibers, while NCK1 binds at Tyr-751 and mediates Cdc42 signaling for filopodium formation. Cells from NCK2-knockout mice and NCK2-knockdown human fibroblasts fail to migrate in response to PDGF-BB. The middle SH3 domain of NCK2 alone (dominant negative) blocks PDGF-BB-induced migration. Knockout MEFs, RNAi knockdown, site-directed mutagenesis of PDGFR-β, dominant-negative SH3 domain overexpression, Rho/Rac/Cdc42 activity assays The Journal of investigative dermatology High 19242519
2010 NCK2 is utilized by the integrin adaptor protein Cas (downstream of integrin signaling) to activate Cdc42 and induce cell polarization during wound healing. This is distinct from CrkII (also recruited by Cas) which activates Rac1 to promote cell protrusion extension. siRNA knockdown, Cdc42/Rac1 activity assays, wound healing assay The FEBS journal Medium 20637038
2011 NCK2 (Nck2) interacts with the SH2 domain of NCK2 via the N-terminus of SKAP2. The SKAP2-NCK2-F-actin complex accumulates at the leading edge of lamellipodia where FGF receptors and focal adhesions are also recruited. NCK2 participates in actin reorganization during lens epithelial cell differentiation. Co-IP, immunofluorescence co-localization, RNAi knockdown, overexpression with deletion mutants Journal of cellular and molecular medicine Medium 20219016
2011 PAK3 interacts preferentially with NCK2 (over NCK1) in brain extracts and transfected cells, independent of PAK3 kinase activity. Selective uncoupling of NCK2 interactions using an interfering peptide in acute cortical slices causes rapid increase in evoked transmission. The PAK3-P12A mutation strongly reduces binding to NCK2 but only slightly to NCK1. Wild-type PAK3 decreases amplitude of spontaneous miniature excitatory currents, whereas P12A mutant does not, demonstrating that PAK3 down-regulates synaptic transmission through its interaction with NCK2. Co-IP from brain extracts and transfected cells, interfering peptide, electrophysiology in cortical slices, mutagenesis The Journal of biological chemistry High 21949127
2011 NCK2, but not NCK1, is required for Slit1-induced changes in cortical neuron morphology in vitro. NCK1 and NCK2 both bind to the Slit receptor Robo via an atypical SH3-mediated mechanism. shRNA knockdown (isoform-specific), Co-IP, morphological analysis of cortical neurons, Robo1/Robo2 knockout neurons Molecular and cellular neurosciences Medium 21600986
2013 NCK1, but not NCK2, is a substrate of c-Cbl-mediated ubiquitination at Lys178. Synaptopodin competes with c-Cbl for binding to NCK1, preventing NCK1 ubiquitination. Expression of c-Cbl-resistant NCK1(K178R) or NCK2 containing the SH3 domain 2 of NCK1 restores stress fibers in synaptopodin-depleted podocytes through RhoA signaling activation. NCK2 is not ubiquitinated by c-Cbl (negative finding that is mechanistically informative—NCK2 lacks the ubiquitin acceptor site equivalent, explaining its differential regulation). Ubiquitination assay, mutagenesis (K178R), c-Cbl knockdown, RhoA activity assay, domain-swap constructs Nature communications High 24287595
2014 In human Jurkat T cells, NCK2 knockdown (to ~10%) reduces TCR-induced NFAT activation but does not significantly impair Erk/MEK phosphorylation, AP-1 activation, or IL-2/CD69 expression (in contrast to NCK1 knockdown). Thus NCK2 has a role in NFAT activation downstream of TCR but not in Erk pathway activation. shRNA knockdown (isoform-specific), luciferase reporter assays, phospho-immunoblotting Cell communication and signaling Medium 24670066
2016 NCK2 deficiency in mice promotes adiposity with adipocyte hypertrophy and enhanced adipogenesis. Mechanistically, NCK2 deficiency in adipocyte precursors is associated with primed PERK activation and signaling, which promotes the adipogenic program including enhanced adipocyte differentiation, lipid droplet formation, and dysfunctional lipogenesis/lipolysis. Knockout mice, in vitro differentiation assays (3T3-L1, stromal vascular fraction), PERK signaling analysis Diabetes Medium 27325288
2018 NCK2 is required for cell abscission during cytokinesis. Nck2-/- (but not Nck1-/-) MEFs are multinucleated, display extended intercellular bridge protrusions, spend extended time in cytokinesis, and fail to complete abscission. The midbody in NCK2-deficient cells is longer and shows mislocalization of AURKB, PLK1, and ECT2. NCK2's SH2 domain is required for its cytokinesis function. AP-MS and BioID identified 28 proteins specifically associated with NCK2 (not NCK1). Knockout MEFs, AP-MS, BioID proximity labeling, live imaging, immunofluorescence, domain-swap/SH2 mutant rescue Molecular & cellular proteomics High 30002203
2018 NCK2 is required for Eph-mediated motor axon trajectory selection in spinal lateral motor column neurons. NCK2 loss- and gain-of-function (via in ovo electroporation) perturbs LMC axon trajectory selection and growth preference against ephrins. NCK2 modulates α2-chimaerin activity in the context of Eph signaling. In ovo electroporation (loss- and gain-of-function), in vitro neurite growth assay with ephrins, epistasis with α2-chimaerin Developmental dynamics Medium 30016580
2020 NCK1 (not NCK2) mediates disturbed flow-induced endothelial permeability and PAK2 activation. Both NCK1 and NCK2 bind PECAM-1 in an SH2-dependent manner in response to shear stress, but only NCK1 ablation interferes with PAK2 membrane translocation and activation. Domain-swap experiments show NCK1 SH3 domains (specifically the first SH3) are critical for PAK recruitment and activation. NCK2 depletion has no significant effect on permeability (negative finding). Knockout cells, domain-swap constructs, in vivo partial carotid ligation, PAK activity assay, permeability assays (Evans blue, FITC-dextran), single-point SH3 mutations Journal of the American Heart Association High 32468886
2022 CBL (Casitas B lymphoma E3 ubiquitin ligase) interacts with NCK2, promotes its ubiquitin-mediated proteasomal degradation, and thereby reduces NCK2 protein stability in breast cancer cells. NCK2 overexpression reverses CBL-mediated inhibition of cell proliferation and migration. Co-IP, IP-mass spectrometry, immunofluorescence co-localization, cycloheximide chase, ubiquitination assay, rescue overexpression Nan fang yi ke da xue xue bao Medium 36504051
2023 OIP5 interacts with NCK2 in human spermatogonial stem cells (SSCs), as demonstrated by Co-IP, IP-MS, and GST pulldown. NCK2 silencing decreases SSC proliferation and DNA synthesis but enhances apoptosis. NCK2 knockdown reverses the influence of OIP5 overexpression on SSC self-renewal. Co-IP, IP-mass spectrometry, GST pulldown, siRNA knockdown, proliferation and apoptosis assays Research (Washington, D.C.) Medium 37292517
2024 NCK2 is associated with EphA4 and RhoA in the olfactory bulb (demonstrated by immunoprecipitation), functioning as a scaffolding protein to modulate EphA4/RhoA pathway. α-synuclein deletion reduces NCK2 levels and EphA4 activation; re-expressing α-synuclein reverses NCK2 downregulation and restores pEphA4 and RhoA activity, improving olfactory neuron projection. α-synuclein knockout mice, immunoprecipitation, iTRAQ-LC-MS, overexpression rescue Cell & bioscience Medium 39543759
2024 KLF4 transcription factor binds directly to the NCK2 promoter and facilitates NCK2 transcription, suppressing antitumor effects of brusatol in melanoma. The miR-150-3p/KLF4/NCK2 axis regulates brusatol's anti-melanoma activity. ChIP, promoter reporter assay, overexpression and knockdown, Western blot Biochemical pharmacology Medium 38583810
2025 NCK2 (and NCK1) are enriched within clathrin-coated pits (CCPs) at the plasma membrane and regulate early CCP formation and maturation. NCK2 supports EGF-stimulated Akt phosphorylation. EGF stimulation triggers NCK-dependent enrichment of PI3K within CCPs. Perturbation of NCK adaptors suppresses cell proliferation and survival. TIRFM (total internal reflection fluorescence microscopy), CCP dynamics analysis, Akt phosphorylation assay, PI3K localization bioRxivpreprint Medium bio_10.1101_2025.10.24.684344

Source papers

Stage 0 corpus · 42 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1998 Nck-2, a novel Src homology2/3-containing adaptor protein that interacts with the LIM-only protein PINCH and components of growth factor receptor kinase-signaling pathways. Molecular biology of the cell 166 9843575
2006 The phosphotyrosine peptide binding specificity of Nck1 and Nck2 Src homology 2 domains. The Journal of biological chemistry 83 16636066
2000 Nckbeta adapter regulates actin polymerization in NIH 3T3 fibroblasts in response to platelet-derived growth factor bb. Molecular and cellular biology 75 11027258
1999 Identification of Grb4/Nckbeta, a src homology 2 and 3 domain-containing adapter protein having similar binding and biological properties to Nck. The Journal of biological chemistry 65 10026169
2013 Proteasomal degradation of Nck1 but not Nck2 regulates RhoA activation and actin dynamics. Nature communications 51 24287595
2001 Identification and kinetic analysis of the interaction between Nck-2 and DOCK180. FEBS letters 39 11240126
2006 Structural insight into the binding diversity between the human Nck2 SH3 domains and proline-rich proteins. Biochemistry 37 16752908
2011 Nck2 promotes human melanoma cell proliferation, migration and invasion in vitro and primary melanoma-derived tumor growth in vivo. BMC cancer 36 21992144
2002 Nck-2 interacts with focal adhesion kinase and modulates cell motility. The international journal of biochemistry & cell biology 34 11950595
2011 p21-Activated kinase 3 (PAK3) protein regulates synaptic transmission through its interaction with the Nck2/Grb4 protein adaptor. The Journal of biological chemistry 28 21949127
2011 SKAP2, a novel target of HSF4b, associates with NCK2/F-actin at membrane ruffles and regulates actin reorganization in lens cell. Journal of cellular and molecular medicine 27 20219016
2014 Non-overlapping functions of Nck1 and Nck2 adaptor proteins in T cell activation. Cell communication and signaling : CCS 26 24670066
2007 Nckbeta adapter controls neuritogenesis by maintaining the cellular paxillin level. Molecular and cellular biology 26 17591694
2018 Proteomic Analysis of NCK1/2 Adaptors Uncovers Paralog-specific Interactions That Reveal a New Role for NCK2 in Cell Abscission During Cytokinesis. Molecular & cellular proteomics : MCP 22 30002203
2013 Association of HK2 and NCK2 with normal tension glaucoma in the Japanese population. PloS one 21 23349798
2016 Nck2 Deficiency in Mice Results in Increased Adiposity Associated With Adipocyte Hypertrophy and Enhanced Adipogenesis. Diabetes 20 27325288
2009 Non-compensating roles between Nckalpha and Nckbeta in PDGF-BB signaling to promote human dermal fibroblast migration. The Journal of investigative dermatology 20 19242519
2005 Structural insight into the binding diversity between the Tyr-phosphorylated human ephrinBs and Nck2 SH2 domain. The Journal of biological chemistry 19 15764601
2023 OIP5 Interacts with NCK2 to Mediate Human Spermatogonial Stem Cell Self-Renewal and Apoptosis through Cell Cyclins and Cycle Progression and Its Abnormality Is Correlated with Male Infertility. Research (Washington, D.C.) 17 37292517
2001 Src homology 3 domain-dependent interaction of Nck-2 with insulin receptor substrate-1. The Biochemical journal 17 11171109
2010 Cas utilizes Nck2 to activate Cdc42 and regulate cell polarization during cell migration in response to wound healing. The FEBS journal 16 20637038
2008 Microsatellite analysis of the GLC1B locus on chromosome 2 points to NCK2 as a new candidate gene for normal tension glaucoma. The British journal of ophthalmology 16 18723748
2011 The adaptor protein Nck2 mediates Slit1-induced changes in cortical neuron morphology. Molecular and cellular neurosciences 15 21600986
2024 KLF4 suppresses anticancer effects of brusatol via transcriptional upregulating NCK2 expression in melanoma. Biochemical pharmacology 14 38583810
2013 OSU-03012 sensitizes breast cancers to lapatinib-induced cell killing: a role for Nck1 but not Nck2. BMC cancer 14 23706161
2020 Nck1, But Not Nck2, Mediates Disturbed Flow-Induced p21-Activated Kinase Activation and Endothelial Permeability. Journal of the American Heart Association 13 32468886
2002 Brain-derived neurotrophic factor promotes interaction of the Nck2 adaptor protein with the TrkB tyrosine kinase receptor. Biochemical and biophysical research communications 13 12074588
2013 NCK2 is significantly associated with opiates addiction in African-origin men. TheScientificWorldJournal 11 23533358
2015 Targeting of the EGFR/β1 integrin connecting proteins PINCH1 and Nck2 radiosensitizes three-dimensional SCC cell cultures. Oncology reports 8 26004008
2018 Non-catalytic region of tyrosine kinase adaptor protein 2 (NCK2) pathways as factor promoting aggressiveness in ovarian cancer. The International journal of biological markers 7 29218693
2001 Grb4/Nckbeta acts as a nuclear repressor of v-Abl-induced transcription from c-jun/c-fos promoter elements. The Journal of biological chemistry 6 11514578
2018 Nck2 is essential for limb trajectory selection by spinal motor axons. Developmental dynamics : an official publication of the American Association of Anatomists 5 30016580
2018 Aristolochic acid inhibits Slit2-induced migration and tube formation via inactivation of Robo1/Robo2-NCK1/NCK2 signaling pathway in human umbilical vein endothelial cells. Toxicology letters 5 30381256
2017 Nck2, an unexpected regulator of adipogenesis. Adipocyte 5 28425845
2024 Mechanism of action of vinegared Cornu Cervi Degelatinatum in suppressing spleen kidney yang deficient ulcerative colitis through NCK2-JNK pathway. Heliyon 4 38312676
2023 Distinct Requirements for Adaptor Proteins NCK1 and NCK2 in Mammary Gland Development. Journal of mammary gland biology and neoplasia 4 37479911
2019 Association of HK2 and NCK2 with normal-tension glaucoma in a population from the Republic of Korea. Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie 3 31512042
2022 [CBL inhibits proliferation and invasion of breast cancer cells by ubiquitylation-mediated degradation of NCK2]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University 2 36504051
2024 Adapting to change: resolving the dynamic and dual roles of NCK1 and NCK2. The Biochemical journal 1 39392452
2020 A Case of Lung Adenocarcinoma Harboring a Rare LOC285000-ALK-NCK2 Gene Fusion Identified by Next-Generation Sequencing With Long-Term Response to Crizotinib. JTO clinical and research reports 1 34589983
2024 A novel function for α-synuclein as a regulator of NCK2 in olfactory bulb: implications for its role in olfaction. Cell & bioscience 0 39543759
2022 Endothelial NCK2 promotes atherosclerosis progression in male but not female Nck1-null atheroprone mice. Frontiers in cardiovascular medicine 0 36035930

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