Affinage

ZAP70

Tyrosine-protein kinase ZAP-70 · UniProt P43403

Length
619 aa
Mass
69.9 kDa
Annotated
2026-06-11
100 papers in source corpus 47 papers cited in narrative 45 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ZAP-70 is a cytoplasmic Syk-family protein tyrosine kinase that initiates T cell receptor signaling by coupling antigen recognition to downstream Ca2+, Ras/MAPK, and PI3K cascades essential for thymic selection and T cell activation (PMID:8124727, PMID:8202712, PMID:8202713, PMID:7630421). Following TCR stimulation it associates through its tandem SH2 domains with tyrosine-phosphorylated ITAMs on the TCR zeta chain and redistributes from cytosol to the plasma membrane, in a manner requiring Src-family kinase (Lck) activity (PMID:1423621, PMID:7600293, PMID:9813084). Lck phosphorylation of Y493 in the activation loop is required to switch on ZAP-70 catalytic activity, while the SH2-kinase interdomain B region (Y292, Y315, Y319) imposes autoinhibition: crystal structures show Y315/Y319 engage the kinase domain in aromatic contacts incompatible with ITAM binding, so ITAM engagement destabilizes the autoinhibited conformation as the first step of activation (~5-fold), with Lck phosphorylation providing the dominant (~100-fold) activation (PMID:7781602, PMID:17512407, PMID:23530057, PMID:26237552). Once active, ZAP-70 phosphorylates the adaptors LAT and SLP-76—the latter at Y113/Y128 to enable Vav recruitment—nucleating the LAT signalosome that recruits Grb2, PLCγ1, and PI3K p85, with Lck acting as a bridge between phospho-ZAP-70 and LAT to promote efficient LAT phosphorylation (PMID:8702662, PMID:9047237, PMID:9489702, PMID:29915297). ZAP-70 is recruited to and cycles at the immunological synapse, a process dependent on RhoH and ezrin and required for synapse maturation, centrosome and granule polarization, and integrin/actin control, the latter partly kinase-independent (PMID:18025306, PMID:21103055, PMID:24596147, PMID:27869819). Its activity is negatively regulated by the phosphatase SHP-1, by Sts-1/Sts-2, and by ubiquitin-dependent degradation, and positively sustained by the deubiquitinase Otud7b and by TCR-dependent S-acylation that licenses substrate engagement (PMID:8638162, PMID:14738763, PMID:11493682, PMID:26903241, PMID:33482200). Loss-of-function ZAP-70 mutations cause a human SCID lacking CD8+ T cells with non-functional CD4+ T cells, and a murine SH2-domain point mutation that alters TCR signaling thresholds drives autoimmune arthritis (PMID:8124727, PMID:8202712, PMID:8202713, PMID:14647385). ZAP-70 also functions in B cells, contributing to the pro-B/pre-B transition and, when aberrantly expressed, acting as an adaptor that subverts BCR negative selection in CLL and B-ALL (PMID:12705855, PMID:18048647, PMID:33878293).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1992 High

    Established the molecular identity of the kinase physically coupled to the activated TCR, defining the entry point for TCR signal transduction.

    Evidence cDNA cloning and co-immunoprecipitation showing a 70 kDa kinase associating with TCR zeta after stimulation, dependent on Src-family kinases

    PMID:1423621

    Open questions at the time
    • Did not define the structural basis of zeta association
    • Did not identify downstream substrates
  2. 1994 High

    Demonstrated that ZAP-70 is non-redundantly required for human T cell function and development, linking the kinase to a defined immunodeficiency.

    Evidence Human patient genetics, kinase assays, and T cell functional assays revealing SCID with absent CD8+ T cells and non-functional CD4+ cells; reciprocal co-IP in murine thymocytes confirming SH2-mediated zeta association

    PMID:7600293 PMID:8124727 PMID:8202712 PMID:8202713

    Open questions at the time
    • Mechanism of kinase activation not yet defined
    • Substrates downstream of ZAP-70 unknown
  3. 1995 High

    Defined the activation switch and developmental requirement, showing Lck phosphorylation of Y493 turns on catalysis and that ZAP-70 is needed for both positive and negative thymic selection.

    Evidence Baculovirus reconstitution with Lck and site-directed mutagenesis (Y493), plus ZAP-70 knockout mice with human rescue

    PMID:7561679 PMID:7630421 PMID:7781602

    Open questions at the time
    • Structural basis of activation-loop control unresolved
    • Relationship between Y493 phosphorylation and autoinhibition not yet mapped
  4. 1996 High

    Identified physiological substrates and a key negative regulator, transforming ZAP-70 from a receptor-associated kinase into a defined node coupling the TCR to Ras and calcium pathways.

    Evidence In vitro kinase assays with dominant-negative ZAP-70 (SLP-76 substrate), direct SHP-1 binding/phosphatase assays, c-Cbl association, and mutagenesis defining Y292/Y492 as negative-regulatory sites

    PMID:8551236 PMID:8638162 PMID:8663155 PMID:8702662 PMID:8756661 PMID:8943331

    Open questions at the time
    • Mechanism by which interdomain B tyrosines restrain activity not structurally defined
    • Failure to phosphorylate ITAM peptides in vitro left in vivo substrate context unclear
  5. 1997 High

    Resolved the adaptor-coupling logic and membrane-recruitment role, showing ZAP-70 phosphorylates SLP-76 at specific Vav-binding sites and that SH2 domains serve recruitment rather than catalytic activation.

    Evidence In vitro phosphorylation with site mapping (Y113/Y128) and SH2 binding assays; synthetic ligand-induced membrane recruitment with reporter readouts

    PMID:9047237 PMID:9312021

    Open questions at the time
    • How membrane orientation dictates signaling specificity not mechanistically explained
  6. 1998 High

    Identified LAT as the central ZAP-70 substrate nucleating the signalosome and revealed dynamic membrane redistribution plus a c-Cbl regulatory loop.

    Evidence LAT cloning with kinase and dominant-negative assays; GFP-ZAP-70 live imaging/FRAP; c-Cbl knockout mice with ZAP-70 phosphorylation readout; dominant-negative migration/invasion assays

    PMID:9489702 PMID:9671496 PMID:9732296 PMID:9813084

    Open questions at the time
    • Spatial coordination of LAT phosphorylation at membrane not resolved
    • Migration role mechanistically incomplete
  7. 1999 High

    Refined the autoinhibitory and phosphatase-regulatory architecture, establishing interdomain B as an RTK-juxtamembrane-like brake and ZAP-70 as a direct SHP-1 substrate in cells.

    Evidence Deletion mutagenesis with in vitro kinase and pathway reporters; constitutively active SHP-1 mutants in intact T cells

    PMID:10458769 PMID:9858619

    Open questions at the time
    • Atomic basis of interdomain B autoinhibition still inferred, not visualized
  8. 2003 High

    Connected quantitative ZAP-70 signaling thresholds to disease, showing an SH2-domain point mutation reshapes thymic selection to permit autoimmune T cells, and extended ZAP-70 function into B cell development.

    Evidence Spontaneous W163C mouse mutant with thymic selection analysis; Syk/ZAP-70 double-knockout B cell developmental and allelic-exclusion analysis

    PMID:12705855 PMID:14647385

    Open questions at the time
    • How altered signaling thresholds translate to selection of specific autoreactive specificities unresolved
  9. 2007 High

    Provided the structural foundation for autoinhibition and dissected division of labor with Syk during thymocyte development, while extending roles to synapse recruitment and pathogen exploitation.

    Evidence Crystal structure of autoinhibited ZAP-70 defining Y315/Y319 linker contacts; Syk vs ZAP-70 stage-specific knockout epistasis; ezrin interaction with IS recruitment assays; HIV virological synapse loss-of-function

    PMID:17215865 PMID:17512407 PMID:17606633 PMID:18025306

    Open questions at the time
    • Conformational transition from autoinhibited to active state not yet quantified
    • Mechanism of ezrin-mediated recruitment partial
  10. 2014 High

    Quantified the conformational activation hierarchy and separated catalytic from structural functions, while defining how cumulative ZAP-70 signaling integrates over time to dictate selection outcomes.

    Evidence Crystal structure with unmodified Y315/Y319 plus in vitro kinase quantification (~5-fold ITAM vs ~100-fold Lck); chemical-genetic catalytic inhibition in CTLs and synchronized thymic selection models

    PMID:23530057 PMID:24596147 PMID:24908390

    Open questions at the time
    • How kinase-independent integrin/actin function is structurally executed unresolved
  11. 2016 High

    Established the catalytic 'catch-and-release' cycle and the deubiquitination switch that protects active ZAP-70 from phosphatase-mediated inactivation.

    Evidence Live-cell kinase-activity imaging of ZAP-70 cycling at TCRs; Otud7b knockout mice with co-IP and ubiquitination assays showing deubiquitination blocks Sts1/Sts2 association

    PMID:26903241 PMID:27869819

    Open questions at the time
    • Identity of the E3 ligase opposing Otud7b not defined in the corpus
  12. 2018 High

    Defined the bridging mechanism by which Lck scaffolds ZAP-70 to its substrate LAT, explaining efficient signalosome assembly.

    Evidence Domain-specific SH2/SH3 binding studies and LAT proline-rich motif mutagenesis with T cell development assays

    PMID:29915297

    Open questions at the time
    • Stoichiometry and spatial regulation of the Lck-ZAP-70-LAT bridge at the membrane not resolved
  13. 2021 High

    Identified a lipidation modification controlling substrate engagement and clarified the adaptor/oncogenic role of aberrant ZAP-70 in B cell malignancy.

    Evidence Acyl-Biotin Exchange with lipidation-deficient mutant and TCR signaling assays; conditional Zap70 mouse models of B-ALL/B-CLL with BCR pathway analysis; CLL structure-function showing kinase-independent SH2-dependent BCR enhancement

    PMID:18048647 PMID:33482200 PMID:33878293

    Open questions at the time
    • Enzyme catalyzing ZAP-70 S-acylation unidentified
    • How ZAP-70 redirects SYK signaling mechanistically incomplete

Open questions

Synthesis pass · forward-looking unresolved questions
  • The enzymes that S-acylate ZAP-70 and the E3 ligase counteracting Otud7b remain unidentified, and how the membrane-orientation, lipidation, and bridging mechanisms jointly specify substrate selection in space is unresolved.
  • No identified acyltransferase for ZAP-70 lipidation
  • E3 ligase driving ZAP-70 ubiquitination not defined in corpus
  • Integration of conformational, lipid, and scaffold cues into substrate choice unresolved

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 5 GO:0016740 transferase activity 2 GO:0060089 molecular transducer activity 2 GO:0060090 molecular adaptor activity 2 GO:0140657 ATP-dependent activity 1
Localization
GO:0005886 plasma membrane 2 GO:0005634 nucleus 1 GO:0005829 cytosol 1
Pathway
R-HSA-1266738 Developmental Biology 3 R-HSA-162582 Signal Transduction 3 R-HSA-168256 Immune System 3
Partners
C-CBLCD247LATLCKOTUD7BSHP-1SLP-76VAV
Complex memberships
LAT signalosomeTCR/CD3/zeta compleximmunological synapse

Evidence

Reading pass · 45 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1992 ZAP-70 is a 70 kDa protein tyrosine kinase that associates with the TCR zeta chain following T cell receptor stimulation. Tyrosine phosphorylation and association of ZAP-70 with zeta require the presence of src family PTKs. cDNA cloning, co-immunoprecipitation, tyrosine phosphorylation assays Cell High 1423621
1994 ZAP-70 kinase activity is essential for TCR signal transduction and CD8+ T cell development; loss-of-function mutations in ZAP-70 cause human SCID characterized by absence of CD8+ T cells and non-functional CD4+ T cells that fail to produce IL-2 or proliferate in response to TCR stimulation. Human patient genetic analysis, kinase activity assays, T cell functional assays Cell High 8124727 8202712 8202713
1994 ZAP-70 constitutively associates with tyrosine-phosphorylated TCR zeta in murine thymocytes and lymph node T cells via its two SH2 domains; TCR ligation promotes a large increase in ZAP-70 tyrosine phosphorylation. Co-immunoprecipitation, immunoblotting, genetic studies with TCR/coreceptor mutants Immunity High 7600293
1994 ZAP-70 associates with the SH2 domain of proto-Vav following TCR stimulation; this interaction requires tyrosine phosphorylation of ZAP-70 and is inhibited by a ZAP-70-specific synthetic tyrosine phosphopeptide, suggesting ZAP-70 may function as a PTK for proto-Vav. Co-immunoprecipitation, SH2 domain pulldown, peptide competition assay The Journal of biological chemistry Medium 7798261
1995 ZAP-70 kinase activity is required for both positive and negative thymic selection; mice lacking ZAP-70 have no CD4 or CD8 single-positive T cells and thymocytes are not deleted by peptide antigens, while NK cell function remains intact. ZAP-70 knockout mice, thymic reconstitution with human ZAP-70, flow cytometry, functional assays Nature High 7630421
1995 Phosphorylation of ZAP-70 at tyrosine 493 by Lck (a src family PTK) is required for activation of ZAP-70 catalytic activity; Y493 mutation alone abrogated Lck-mediated activation, and this phosphorylation is required for TCR-mediated IL-2 secretion. Baculovirus expression, in vitro kinase assay with Lck co-infection, site-directed mutagenesis, IL-2 secretion assay The EMBO journal High 7781602
1995 Syk compensates for the loss of ZAP-70 in ZAP-70-deficient human thymocytes, allowing selection of CD4+ T cells, but not in peripheral T cells; Syk is present at high levels and is tyrosine phosphorylated after TCR stimulation in thymocyte lines from ZAP-70-deficient patients. Patient thymocyte analysis, immunoblotting, tyrosine phosphorylation assays The Journal of experimental medicine Medium 7561679
1995 Tyrosine-phosphorylated ZAP-70 and Syk bind to the SH2 domain of p56lck via an SH2-mediated interaction after CD3 stimulation, enabling recruitment of CD4 to antigen-stimulated TCR/CD3/zeta complexes. Co-immunoprecipitation, phosphopeptide competition, cocapping experiments The Journal of experimental medicine Medium 7539035
1996 SLP-76 is a direct substrate of ZAP-70; ZAP-70 phosphorylates SLP-76 preferentially in vitro and in heterologous cellular systems, and this phosphorylation is diminished in T cells expressing catalytically inactive ZAP-70. SLP-76 phosphorylation by ZAP-70 links the TCR to Ras and calcium pathways. In vitro kinase assay, dominant-negative ZAP-70 in T cells, overexpression of SLP-76 mutants, functional reporter assays The Journal of biological chemistry High 8702662
1996 SHP-1 phosphatase binds directly to ZAP-70 upon T cell activation, resulting in increased SHP-1 phosphatase activity and decreased ZAP-70 kinase activity; SHP-1 acts as a negative regulator of TCR signaling and sets the threshold of T cell activation. Co-immunoprecipitation, phosphatase activity assay, kinase activity assay, dominant-negative SHP-1 expression in T cells Science High 8638162
1996 ZAP-70 associates with c-Cbl protooncogene product (p120) following TCR stimulation; ZAP-70 can cause tyrosine phosphorylation of p120c-cbl dependent on Lck- or FynT-mediated signals. Co-immunoprecipitation, heterologous expression system, immunoblotting The Journal of experimental medicine Medium 8551236
1996 Distinct tyrosine phosphorylation sites in ZAP-70 mediate activation and negative regulation: phosphorylation of Y493 by Src-PTK is required for TCR-mediated activation, while mutations at Y292 or Y492 result in hyperactive TCR phenotypes, indicating these sites negatively regulate ZAP-70. Site-directed mutagenesis, Syk/ZAP-70-deficient lymphocyte reconstitution, calcium and Ras pathway assays Molecular and cellular biology High 8756661
1996 Mutation of Y292 to phenylalanine (292F) or deletion of interdomain B results in a gain-of-function ZAP-70 with enhanced ability to activate lymphocytes, demonstrating that Y292 negatively regulates ZAP-70 function without affecting kinase activity or receptor binding. Site-directed mutagenesis, NF-AT reporter assay in Syk-deficient cells and T cell lines, in vitro kinase assay Molecular and cellular biology High 8943331
1996 Purified recombinant ZAP-70 exhibits high substrate specificity; kinase activity requires cation with preference for Mn2+ over Mg2+, and among tested substrates only alpha-tubulin and cytoplasmic fragment of erythrocyte band 3 are good substrates. ZAP-70 does not phosphorylate TCRzeta ITAMs or short CD3/zeta peptides in vitro. Baculovirus expression, protein purification, in vitro kinase assay with multiple substrates, Km determination The Journal of biological chemistry High 8663155
1997 ZAP-70 phosphorylates SLP-76 at specific sites (Y113 and Y128, YESP motifs) that allow Vav SH2 domain binding; of the three kinases tested (p56lck, p59fyn, ZAP-70), only ZAP-70 phosphorylates SLP-76 to enable Vav-SLP-76 complex formation. In vitro phosphorylation, SH2 domain binding assay, in vivo analysis in T cells Immunity High 9047237
1997 Transient membrane recruitment of ZAP-70 through synthetic ligands leads to rapid ZAP-70 phosphorylation and activation of Ras/MAPK and Ca2+/calcineurin signaling pathways; ZAP-70 SH2 domains function solely in membrane recruitment and not in kinase activation; the orientation of ZAP-70 at the membrane determines downstream signaling specificity. Synthetic ligand-induced membrane recruitment, reporter gene assays, dominant-negative constructs The EMBO journal Medium 9312021
1998 LAT (linker for activation of T cells) is phosphorylated by ZAP-70/Syk protein tyrosine kinases, leading to recruitment of Grb2, phospholipase C-gamma1, and p85 subunit of PI3K; overexpression of a mutant LAT lacking critical tyrosine residues inhibits T cell activation. cDNA cloning, tyrosine phosphorylation assay, dominant-negative overexpression, co-immunoprecipitation Cell High 9489702
1998 ZAP-70 redistributes from cytosol to the plasma membrane upon TCR activation via TCRzeta chain interaction; this redistribution requires TCRzeta and is enhanced by active Lck. ZAP-70 has decreased mobility at the plasma membrane compared to cytosol, and nuclear ZAP-70 is phosphorylated on tyrosine upon TCR stimulation. Live-cell fluorescence imaging with GFP-tagged ZAP-70, FRAP, confocal microscopy in HeLa and T cells The Journal of cell biology High 9813084
1998 c-Cbl-deficient mice show uncoupled ZAP-70 kinase phosphorylation from CD4-mediated Lck activation in thymocytes following CD3epsilon cross-linking, establishing c-Cbl as a negative regulator of ZAP-70 family kinase signaling. c-Cbl knockout mice, immunoblotting for ZAP-70 phosphorylation, CD3 cross-linking assays Molecular and cellular biology Medium 9671496
1998 ZAP-70 is required for LFA-1-dependent T cell migration; dominant-negative ZAP-70 blocks invasion of T cell hybridoma cells into fibroblast monolayers, and LFA-1 cross-linking induces tyrosine phosphorylation blocked by dominant-negative ZAP-70. Dominant-negative ZAP-70 overexpression, invasion assay, in vivo metastasis model, LFA-1 cross-linking with phosphorylation assay The Journal of cell biology Medium 9732296
1999 Constitutively active SHP-1 (lacking SH2 domains) dephosphorylates ZAP-70 and Syk in intact cells and inhibits downstream events including Erk2 activation and IL-2 gene transcription, identifying ZAP-70 as a direct substrate of SHP-1 in T cells. Constitutively active and inactive SHP-1 mutant expression, immunoblotting for ZAP-70/Syk phosphorylation, Erk2 assay, IL-2 reporter assay European journal of immunology Medium 10458769
1999 Interdomain B of ZAP-70 (containing Y292, Y315, Y319) negatively regulates ZAP-70 catalytic activity analogously to juxtamembrane regions of receptor tyrosine kinases, but is not required for ZAP-70 signaling to NF-AT, calcium fluxes, or MAPK activation. Deletion mutagenesis, in vitro kinase assay, NF-AT reporter assay, calcium flux measurement, dominant-negative Ras Molecular and cellular biology High 9858619
2001 ZAP-70 and Syk become ubiquitinated in response to CD16 aggregation on NK cells, and the ubiquitinated forms associate with the receptor complex; proteasome and lysosomal inhibitors counteract ZAP-70/Syk degradation, establishing ubiquitin-dependent proteolysis as a negative regulatory mechanism for these kinases. Co-immunoprecipitation, in vitro ubiquitination assay, proteasome/lysosome inhibitors, immunoblotting Proceedings of the National Academy of Sciences Medium 11493682
2002 VHR (a dual-specific phosphatase) is phosphorylated at Y138 by ZAP-70; this phosphorylation enables VHR to inhibit the Erk2-Elk-1 pathway, and the VHR(Y138F) mutant augments TCR-induced Erk2 kinase activity and IL-2 gene expression, establishing VHR as a ZAP-70 substrate that tempers Erk2 activation. In vitro kinase assay, site-directed mutagenesis, Erk2 kinase assay, IL-2 reporter assay in T cells Nature immunology High 12447358
2002 ZAP-70 expression in CLL B cells is associated with enhanced tyrosine phosphorylation of cytosolic proteins including p72Syk following BCR ligation; ZAP-70 undergoes tyrosine phosphorylation and associates with surface immunoglobulin and CD79b upon BCR stimulation, indicating ZAP-70 participates in BCR signaling. Co-immunoprecipitation, tyrosine phosphorylation assays, BCR ligation experiments, Western blotting Blood Medium 12393534
2002 ZAP-70 is required for CXCL12-mediated T cell transendothelial migration; ZAP-70-deficient or kinase-dead ZAP-70 T cells fail to migrate in response to CXCL12, and CXCL12 induces ZAP-70 phosphorylation; the ZAP-70/Vav1 pathway (but not ERK) is required for chemotaxis. Pharmacologic inhibition, dominant-negative ZAP-70, ZAP-70-deficient patient cells, ZAP-70 transfection rescue, migration assay, phosphorylation assays Blood Medium 11964272
2003 A spontaneous point mutation in an SH2 domain of ZAP-70 (W163C in mice) causes chronic autoimmune arthritis by altering TCR signaling thresholds, leading to positive selection of otherwise negatively selected autoimmune T cells. Mouse genetics, thymic selection assays, T cell receptor signaling analysis Nature High 14647385
2003 ZAP-70 is expressed throughout B cell development and plays a role in the pro-B to pre-B cell transition; mice deficient in both Syk and ZAP-70 show a complete block at the pro-B cell stage and failure of heavy chain allelic exclusion, demonstrating ZAP-70 contributes to pre-BCR signaling. ZAP-70 and Syk single/double-knockout mice, flow cytometry of B cell developmental stages, allelic exclusion analysis Immunity High 12705855
2004 Sts-1 and Sts-2 negatively regulate ZAP-70 by opposing its phosphorylation and activation; T cells from Sts-1/Sts-2 double-knockout mice show increased ZAP-70 phosphorylation and ubiquitinylation, hyperactivation of downstream TCR signaling, and increased cytokine production. Sts-1/Sts-2 double-knockout mice, ZAP-70 phosphorylation assays, cytokine assays, autoimmune model Immunity Medium 14738763
2005 Interdomain B of ZAP-70 downregulates ZAP-70 catalytic activity via an autoinhibitory mechanism analogous to juxtamembrane regions of receptor tyrosine kinases such as EphB2; similar regulation applies to the related Syk kinase. Site-directed mutagenesis of ZAP-70 interdomain B, in vitro kinase assay, comparison with RTK autoinhibition models Molecular and cellular biology Medium 15923611
2007 Crystal structure of autoinhibited ZAP-70 reveals that the inactive kinase domain adopts a conformation similar to CDKs and Src kinases; autoinhibition involves interactions between the regulatory segment and the hinge region of the kinase domain; two tyrosines (Y315, Y319) in the SH2-kinase linker engage in aromatic-aromatic interactions that connect the linker to the kinase domain, inconsistent with ITAM binding, suggesting ITAM engagement destabilizes the autoinhibited conformation as the first step in kinase activation. X-ray crystallography of autoinhibited ZAP-70 variant Cell High 17512407
2007 ZAP-70 kinase activity is required for HIV cell-to-cell spread and virological synapse formation; in ZAP-70-deficient or kinase-dead ZAP-70 lymphocytes, intracellular Gag localization is impaired and HIV transmission to recipient cells is reduced. ZAP-70-deficient T cells, kinase-dead ZAP-70 mutant, HIV replication assay, virological synapse analysis The EMBO journal Medium 17215865
2007 Ezrin directly interacts with ZAP-70 and recruits it to the immunological synapse; phosphorylation-activated ezrin is essential for this recruitment, while moesin dephosphorylation and removal are required for IS formation. Direct interaction assay, immunological synapse formation assay, phospho-specific analysis, siRNA knockdown The Journal of cell biology Medium 18025306
2007 Syk and ZAP-70 provide distinct, temporally separated fitness advantages during early thymocyte development: Syk is specifically required for initial pre-TCR signaling at DN3 beta-selection stage, while ZAP-70 promotes sustained pre-TCR/TCR signaling during DN4, ISP, and DP stages before positive selection. Syk- and ZAP-70-deficient mice, in vivo anti-CD3 treatment of RAG1-deficient mice, flow cytometry, cell-cycle analysis The Journal of experimental medicine High 17606633
2007 ZAP-70 kinase activity is dispensable for ZAP-70 to enhance IgM signaling in CLL B cells; a kinase-defective ZAP-70 mutant and a ZAP-70 unable to bind c-Cbl both enhanced BCR signaling; the SH2 domain (but not kinase domain) was necessary for enhanced calcium flux, establishing ZAP-70 acts as an adaptor protein in CLL BCR signaling. Lentiviral transduction of CLL cells with ZAP-70 mutants, calcium flux assay, phosphorylation assays Blood High 18048647
2010 RhoH regulates TCR signaling by recruiting ZAP-70 and Lck to the immunological synapse; RhoH-deficient T cells show impaired Lck-CD3zeta association and defective translocation of ZAP-70 to the IS; forced membrane-targeted ZAP-70 (myr-ZAP-70) partially rescued thymic development defects in RhoH-/- mice. RhoH knockout mice, co-immunoprecipitation, subcellular fractionation, rescue with myristoylated ZAP-70, flow cytometry PloS one Medium 21103055
2013 Crystal structure of ZAP-70 with unmodified Y315 and Y319 reveals distinct roles: Y315 is part of a hydrophobic interface between the regulatory apparatus and kinase domain (disrupted by ITAM engagement), while Y319 suppresses ZAP-70 activity even after SH2 domain disengagement. ITAM binding activates ZAP-70 ~5-fold while Lck phosphorylation activates it ~100-fold. X-ray crystallography, in vitro kinase assay with ITAM peptide and Lck phosphorylation Molecular and cellular biology High 23530057
2014 Zap70 catalytic activity is required to phosphorylate Vav-1 and complete immunological synapse formation in CTLs; without Zap70 catalytic activity, IS formation is arrested at an actin/integrin-rich interdigitation stage, centrosome polarization aborts, and granules fail to polarize; Zap70 also has a structural (kinase-independent) role in integrin-mediated actin control. Chemical-genetic inhibition of Zap70 catalytic activity, synapse formation assay, cytoskeletal imaging, centrosome polarization assay eLife High 24596147
2014 CD4+ CD8+ thymocytes integrate multiple transient Zap70-dependent signals over >36 hours to reach a cumulative threshold for positive selection, while 1 hour of Zap70 signaling is sufficient for negative selection; titration of Zap70 activity results in graded reductions in both selection processes. Chemical-genetic synchronized thymic selection model with selective Zap70 inhibition, flow cytometry Nature immunology High 24908390
2015 TCR dwell times control Zap70 kinase activity; TCR binding or phosphorylation of Zap70 triggers a transition from closed (autoinhibited) to open conformation; the closed conformation minimizes TCR dwell times preventing activation; parallel recruitment of coreceptor-associated Lck ensures Zap70 phosphorylation and stabilizes TCR binding. Structural analysis of Zap70 conformational states, defined-conformation Zap70 mutants, TCR binding assays Nature immunology High 26237552
2016 Otud7b deubiquitinase is recruited to ZAP-70 upon TCR ligation and deubiquitinates ZAP-70, thereby preventing association of ZAP-70 with the negative-regulatory phosphatases Sts1 and Sts2; Otud7b deficiency attenuates ZAP-70 activation and impairs T cell activation and differentiation. Co-immunoprecipitation, ubiquitination assay, Otud7b knockout mice, T cell activation and differentiation assays The Journal of experimental medicine High 26903241
2016 A cycle of Zap70 recruitment, activation, and release at phosphorylated TCRs amplifies and disperses antigenic stimuli; activated Zap70 released from the TCR remains at the membrane, translocates, and phosphorylates spatially distinct substrates, turning the TCR into a catalytic unit. Live-cell imaging, fluorescence-based kinase activity reporters, TCR phosphorylation assays Nature immunology Medium 27869819
2018 Lck bridges Zap70 to LAT by simultaneously binding phospho-Zap70 via its SH2 domain and a conserved proline-rich motif in LAT via its SH3 domain, facilitating efficient LAT phosphorylation by Zap70; elimination of the LAT proline-rich motif compromises TCR signaling and T cell development. Biochemical interaction assays, SH2/SH3 domain binding studies, LAT proline-rich motif mutagenesis, T cell signaling and development assays Nature immunology High 29915297
2021 ZAP-70 undergoes TCR-dependent S-acylation (lipidation); this post-translational modification is dispensable for enzymatic activity but essential for ZAP-70 interaction with its protein substrates and propagation of the TCR signaling pathway; kinetics of S-acylation are consistent with TCR signaling events. Acyl-Biotin Exchange assay, lipidation-deficient ZAP-70 mutant, TCR signaling assays The Journal of biological chemistry Medium 33482200
2021 Aberrant ZAP-70 expression in B cells competes with SYK at the BCR signalosome and redirects SYK from NFAT-dependent negative selection toward tonic PI3K signaling, promoting B cell survival and subverting negative selection of autoreactive and premalignant clones. Genetic mouse models for B-ALL and B-CLL with conditional Zap70 expression/deletion, BCR signaling pathway analysis, negative selection assays Molecular cell High 33878293

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1998 LAT: the ZAP-70 tyrosine kinase substrate that links T cell receptor to cellular activation. Cell 1007 9489702
2003 ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. The New England journal of medicine 1003 12724482
1992 ZAP-70: a 70 kd protein-tyrosine kinase that associates with the TCR zeta chain. Cell 991 1423621
2004 ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia. The New England journal of medicine 706 15329427
2003 Altered thymic T-cell selection due to a mutation of the ZAP-70 gene causes autoimmune arthritis in mice. Nature 673 14647385
1994 Defective T cell receptor signaling and CD8+ thymic selection in humans lacking zap-70 kinase. Cell 468 8124727
1995 Essential role for ZAP-70 in both positive and negative selection of thymocytes. Nature 456 7630421
1994 Human severe combined immunodeficiency due to a defect in ZAP-70, a T cell tyrosine kinase. Science (New York, N.Y.) 417 8202712
2002 Expression of ZAP-70 is associated with increased B-cell receptor signaling in chronic lymphocytic leukemia. Blood 409 12393534
1994 ZAP-70 deficiency in an autosomal recessive form of severe combined immunodeficiency. Science (New York, N.Y.) 407 8202713
1996 Direct regulation of ZAP-70 by SHP-1 in T cell antigen receptor signaling. Science (New York, N.Y.) 348 8638162
1995 Activation of ZAP-70 kinase activity by phosphorylation of tyrosine 493 is required for lymphocyte antigen receptor function. The EMBO journal 335 7781602
1996 Phosphorylation of SLP-76 by the ZAP-70 protein-tyrosine kinase is required for T-cell receptor function. The Journal of biological chemistry 312 8702662
1998 Tissue hyperplasia and enhanced T-cell signalling via ZAP-70 in c-Cbl-deficient mice. Molecular and cellular biology 309 9671496
2010 ZAP-70: an essential kinase in T-cell signaling. Cold Spring Harbor perspectives in biology 303 20452964
2008 Relative value of ZAP-70, CD38, and immunoglobulin mutation status in predicting aggressive disease in chronic lymphocytic leukemia. Blood 245 18577710
1994 ZAP-70 is constitutively associated with tyrosine-phosphorylated TCR zeta in murine thymocytes and lymph node T cells. Immunity 229 7600293
2007 Structural basis for the inhibition of tyrosine kinase activity of ZAP-70. Cell 201 17512407
1997 Regulation of Vav-SLP-76 binding by ZAP-70 and its relevance to TCR zeta/CD3 induction of interleukin-2. Immunity 190 9047237
2009 The structure, regulation, and function of ZAP-70. Immunological reviews 164 19290920
2004 Regulation of ZAP-70 activation and TCR signaling by two related proteins, Sts-1 and Sts-2. Immunity 153 14738763
2014 ZAP-70 genotype disrupts the relationship between microbiota and host, leading to spondyloarthritis and ileitis in SKG mice. Arthritis & rheumatology (Hoboken, N.J.) 151 25048686
2014 MIF promotes B cell chemotaxis through the receptors CXCR4 and CD74 and ZAP-70 signaling. Journal of immunology (Baltimore, Md. : 1950) 136 24760155
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1996 SRK, the stigma-specific S locus receptor kinase of Brassica, is targeted to the plasma membrane in transgenic tobacco. The Plant cell 98 8721749
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2004 Immunohistochemical detection of ZAP-70 in 341 cases of non-Hodgkin and Hodgkin lymphoma. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 73 15133473
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2014 Quantitative and temporal requirements revealed for Zap70 catalytic activity during T cell development. Nature immunology 62 24908390
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1998 ZAP-70 association with T cell receptor zeta (TCRzeta): fluorescence imaging of dynamic changes upon cellular stimulation. The Journal of cell biology 55 9813084
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