Asunto(s)
Adhesión Celular/fisiología , Granulocitos/patología , Integrina beta1/fisiología , Janus Quinasa 2/metabolismo , Trastornos Mieloproliferativos/patología , Molécula 1 de Adhesión Celular Vascular/metabolismo , Enfermedad Crónica , Humanos , Trastornos Mieloproliferativos/enzimología , Trastornos Mieloproliferativos/metabolismoAsunto(s)
Enfermedad Injerto contra Huésped/inmunología , Quinasas Janus/antagonistas & inhibidores , Trastornos Mieloproliferativos/inmunología , Inhibidores de Proteínas Quinasas/farmacología , Linfocitos T/inmunología , Animales , Benzamidas/farmacología , Células Cultivadas , Modelos Animales de Enfermedad , Enfermedad Injerto contra Huésped/tratamiento farmacológico , Enfermedad Injerto contra Huésped/metabolismo , Enfermedad Injerto contra Huésped/patología , Humanos , Ratones , Ratones Endogámicos BALB C , Trastornos Mieloproliferativos/tratamiento farmacológico , Trastornos Mieloproliferativos/metabolismo , Trastornos Mieloproliferativos/patología , Pirimidinas/farmacología , Linfocitos T/efectos de los fármacos , Linfocitos T/patologíaRESUMEN
The purpose of this study was the appraisal of the clinical and functional consequences of germline mutations within the gene for the IL-2 inducible T-cell kinase, ITK. Among patients with Epstein-Barr virus-driven lymphoproliferative disorders (EBV-LPD), negative for mutations in SH2D1A and XIAP (n=46), we identified two patients with R29H or D500T,F501L,M503X mutations, respectively. Human wild-type (wt) ITK, but none of the mutants, was able to rescue defective calcium flux in murine Itk(-/-) T cells. Pulse-chase experiments showed that ITK mutations lead to varying reductions of protein half-life from 25 to 69% as compared with wt ITK (107 min). The pleckstrin homology domain of wt ITK binds most prominently to phosphatidylinositol monophosphates (PI(3)P, PI(4)P, PI(5)P) and to lesser extend to its double or triple phosphorylated derivates (PIP2, PIP3), interactions which were dramatically reduced in the patient with the ITK(R29H) mutant. ITK mutations are distributed over the entire protein and include missense, nonsense and indel mutations, reminiscent of the situation in its sister kinase in B cells, Bruton's tyrosine kinase.
Asunto(s)
Mutación de Línea Germinal , Herpesvirus Humano 4/fisiología , Trastornos Linfoproliferativos/virología , Proteínas Tirosina Quinasas/genética , Sitios de Unión , Niño , Preescolar , Femenino , Humanos , Masculino , Mutación Missense , Linaje , Fosforilación , Proteínas Tirosina Quinasas/metabolismoRESUMEN
T lymphocytes are central players in the adaptive immune response to pathogens. Cytotoxic T cells are able to identify and eliminate virally infected cells, while helper T cells support B lymphocyte-dependent antibody production as well as produce the cytokines that will determine whether a cell- or antibody-mediated immune response is required. The activation of T cells by pathogens is a complex process requiring multiple tightly regulated signaling pathways. Defects within this network, however, can cause severe and chronic disorders such as autoimmunity. Therefore, improving our understanding of how T cells discriminate between antigens and how these signals are organized to yield distinct immune responses is of importance as this may lead to the identification of novel drug targets and better therapeutic strategies.
Asunto(s)
Activación de Linfocitos/inmunología , Biología de Sistemas , Linfocitos T/inmunología , Presentación de Antígeno/inmunología , Tolerancia Inmunológica/inmunología , Receptores de Antígenos de Linfocitos T/inmunologíaRESUMEN
SKAP-HOM is a cytosolic adaptor protein representing a specific substrate for the Src family protein tyrosine kinase Fyn. Previously, several groups have provided experimental evidence that SKAP-HOM (most likely in cooperation with the cytosolic adaptor protein ADAP) is involved in regulating leukocyte adhesion. To further assess the physiological role of SKAP-HOM, we investigated the immune system of SKAP-HOM-deficient mice. Our data show that T-cell responses towards a variety of stimuli are unaffected in the absence of SKAP-HOM. Similarly, B-cell receptor (BCR)-mediated total tyrosine phosphorylation and phosphorylation of Erk, p38, and JNK, as well as immunoreceptor-mediated Ca(2+) responses, are normal in SKAP-HOM(-/-) animals. However, despite apparently normal membrane-proximal signaling events, BCR-mediated proliferation is strongly attenuated in the absence of SKAP-HOM(-/-). In addition, adhesion of activated B cells to fibronectin (a ligand for beta1 integrins) as well as to ICAM-1 (a ligand for beta2 integrins) is strongly reduced. In vivo, the loss of SKAP-HOM results in a less severe clinical course of experimental autoimmune encephalomyelitis following immunization of mice with the encephalitogenic peptide of MOG (myelin oligodendrocyte glycoprotein). This is accompanied by strongly reduced serum levels of MOG-specific antibodies and lower MOG-specific T-cell responses. In summary, our data suggest that SKAP-HOM is required for proper activation of the immune system, likely by regulating the cross-talk between immunoreceptors and integrins.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/fisiología , Linfocitos B/inmunología , Fosfoproteínas/fisiología , Proteínas Adaptadoras Transductoras de Señales/análisis , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Adhesión Celular/inmunología , Citosol/química , Citosol/metabolismo , Sistema Hematopoyético/citología , Sistema Hematopoyético/metabolismo , Inmunoglobulinas/sangre , Integrinas/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Ratones , Ratones Mutantes , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas de la Mielina , Glicoproteína Asociada a Mielina/inmunología , Glicoproteína Mielina-Oligodendrócito , Fosfoproteínas/análisis , Fosfoproteínas/genética , Receptores de Antígenos de Linfocitos B/metabolismo , Transducción de SeñalRESUMEN
AIMS: Due to the systemic character of Type 2 diabetes, cellular disturbances paralleled by an altered expression of various growth factors constitute the basis for impaired wound healing. Cell-surface antigens are altered in chronic wounds and may also have an effect on the persistance of diabetic foot lesions. METHODS: We investigated blood samples of diabetic patients with diabetic foot ulcers (n=21) in comparison with those from healthy control patients subsequent to an injury (n=9). A blood sample (EDTA) was taken from each participant (in the trauma control group on the third day after injury) and examined by flow cytometry [fluorescence-activated cell sorter (FACS)]. Typical cell surface antigens involved in wound healing were studied [cluster of differentiation (CD)2, CD3, CD4, CD25 and human leukocyte antigen (HLA)-diabetic retinopathy (DR)]. RESULTS: known to adversely affect wound healing were elevated in diabetic patients (CD2 p<0.001; CD3 p=0.016, CD4 p=0.22, CD25 p<0.001). HLA-DR expression was also decreased in diabetic foot patients (p=0.023). CONCLUSIONS: Cell-surface antigens appear to be altered in diabetic patients when compared to healthy controls. Thus, due to the systemic character of Type 2 diabetes, cellular disturbances may well constitute the basis for impaired wound healing in diabetes.
Asunto(s)
Antígenos de Diferenciación de Linfocitos B/sangre , Antígenos de Diferenciación de Linfocitos T/sangre , Diabetes Mellitus Tipo 2/inmunología , Pie Diabético/inmunología , Cicatrización de Heridas/inmunología , Adulto , Antígenos de Diferenciación de Linfocitos B/inmunología , Antígenos de Diferenciación de Linfocitos T/inmunología , Antígenos CD2/sangre , Complejo CD3/sangre , Antígenos CD4/sangre , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/complicaciones , Pie Diabético/patología , Femenino , Citometría de Flujo , Antígenos HLA-DR/sangre , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Receptores de Interleucina-2/sangre , Heridas y Lesiones/inmunologíaAsunto(s)
Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/genética , Cromosomas Humanos Par 9/genética , Proteínas de la Membrana/genética , Regiones Promotoras Genéticas , Secuencia de Bases , Sitios de Unión , Mapeo Cromosómico , Cromosomas Artificiales , Exones , Expresión Génica , Humanos , Hibridación Fluorescente in Situ , Intrones , Tejido Linfoide , Glicoproteínas de Membrana , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Transducción de Señal , Factores de TranscripciónRESUMEN
SIT (SHP2-interacting transmembrane adaptor protein) is a recently identified transmembrane adaptor protein, which is expressed in lymphocytes. Its structural properties, in particular the presence of five potential tyrosine phosphorylation sites, suggest involvement of SIT in TCR-mediated recruitment of SH2 domain-containing intracellular signaling molecules to the plasma membrane. Indeed, it has recently been demonstrated that SIT inducibly interacts with the SH2-containing protein tyrosine phosphatase 2 (SHP2) via an immunoreceptor tyrosine-based inhibition motif (ITIM). Moreover, SIT is capable to inhibit TCR-mediated signals proximal of activation of protein kinase C. However, inhibition of T cell activation by SIT occurs independently of SHP2 binding. The present study was performed to further characterize the molecular interaction between SIT and intracellular effector molecules and to identify the protein(s) mediating its inhibitory function. We demonstrate that SIT not only interacts with SHP2 but also with the adaptor protein Grb2 via two consensus YxN motifs. However, mutation of both Grb2-binding sites also does not influence the inhibitory function of SIT. In contrast, mutation of the tyrosine-based signaling motif Y(168) ASV completely abrogates the ability of SIT to inhibit T cell activation. Co-precipitation experiments revealed that the tyrosine kinase p50(csk) could represent the negative regulatory effector molecule that binds to this motif.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas Tirosina Fosfatasas/metabolismo , Linfocitos T/inmunología , Secuencia de Aminoácidos , Sitios de Unión , Proteínas Portadoras/genética , Citoplasma/metabolismo , Proteína Adaptadora GRB2 , Humanos , Péptidos y Proteínas de Señalización Intracelular , Células Jurkat , Activación de Linfocitos , Glicoproteínas de Membrana/genética , Proteínas de la Membrana/genética , Datos de Secuencia Molecular , Fosforilación , Proteína Fosfatasa 2 , Estructura Terciaria de Proteína , Proteína Tirosina Fosfatasa no Receptora Tipo 11 , Proteína Tirosina Fosfatasa no Receptora Tipo 6 , Proteínas Tirosina Fosfatasas/química , Proteínas/metabolismo , Proteínas Tirosina Fosfatasas con Dominio SH2 , Linfocitos T/metabolismo , Tirosina/metabolismoRESUMEN
TRIM is a recently identified transmembrane adaptor protein which is exclusively expressed in T cells and natural killer (NK) cells. In peripheral blood T cells TRIM has been reported to coprecipitate, comodulate, and cocap with the T-cell receptor (TCR), suggesting that it is an integral component of the TCR/CD3/zeta complex. Here we investigate the expression of TRIM mRNAs and proteins in developing thymocytes. Two splicing isoforms with open reading frames are observed, namely a full length (TRIM) and a truncated version (DeltaTM-TRIM). The latter lacks the extracellular and transmembrane domains as well as the first 10 cytoplasmic aminoacids and is significantly expressed only as mRNA in early fetal thymocytes. TRIM mRNA is detected in all mainstream thymocyte subsets in adult mice. TRIM protein, in contrast, first appears in the DN2 (CD44+ CD25+) subset of adult double negative (DN) cells. In fetal thymocyte development, TRIM mRNA is seen from dg 14.5 onwards whereas TRIM protein appears first on dg 16.5. In contrast to the adult, the TRIM protein was seen in a subset of fetal DN1 cells. In fetal and adult thymocytes, TRIM protein expression was highest in DN2, DN3 (CD44-25+) and in DP cells, compatible with a functional role at or around phases of thymic selection.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas de la Membrana/genética , Linfocitos T/metabolismo , Animales , Secuencia de Bases , Proteínas Portadoras/biosíntesis , ADN Complementario , Proteínas de la Membrana/biosíntesis , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , ARN Mensajero , Subgrupos de Linfocitos T/metabolismoRESUMEN
Adapter proteins are well recognised as important molecular switches connecting immunoreceptors with intracellular signalling pathways. However, recent data suggest that homeostasis within the lymphatic system also depends on the coordinated activities of negative regulatory adapter proteins. These prevent activation of lymphocytes in the absence of externally applied signals and regulate termination/limitation of ongoing immune responses via different mechanisms.
Asunto(s)
Linfocitos B/inmunología , Proteínas Portadoras/fisiología , Fosfoproteínas/fisiología , Transducción de Señal/inmunología , Linfocitos T/inmunología , Animales , Linfocitos B/metabolismo , Comunicación Celular/inmunología , Humanos , Activación de Linfocitos/inmunología , Linfocitos T/metabolismoRESUMEN
Protein kinase C theta (PKC theta) is known to induce NF-kappa B, an essential transcriptional element in T cell receptor/CD28-mediated interleukin-2 production but also T cell survival. Here we provide evidence that PKC theta is physically and functionally coupled to Akt1 in this signaling pathway. First, T cell receptor/CD3 ligation was sufficient to induce activation as well as plasma membrane recruitment of PKC theta. Second, PKC theta selectively cooperated with Akt1, known to act downstream of CD28 co-receptor signaling, in activating a NF-kappa B reporter in T cells. Third, Akt1 function was shown to be required for PKC theta-mediated NF-kappa B transactivation. Fourth, PKC theta co-immunoprecipitated with Akt1; however, neither Akt1 nor PKC theta served as a prominent substrate for each other in vitro as well as in intact T cells. Finally, plasma membrane targeting of PKC theta and Akt1 exerted synergistic transactivation of the I-kappa B kinase beta/inhibitor of NF-kappa B/NF-kappa B signaling cascade independent of T cell activation. Taken together, these findings suggest a direct cross-talk between PKC theta and Akt1 in Jurkat T cells.
Asunto(s)
Isoenzimas/metabolismo , FN-kappa B/metabolismo , Proteína Quinasa C/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas , Activación Transcripcional , Secuencia de Bases , Catálisis , Cartilla de ADN , Activación Enzimática , Humanos , Isoenzimas/genética , Células Jurkat , Proteína Quinasa C/genética , Proteína Quinasa C-theta , Proteínas Serina-Treonina Quinasas/genética , Proteínas Proto-Oncogénicas c-akt , Proteínas Recombinantes de Fusión/metabolismo , Transducción de SeñalRESUMEN
In resting peripheral T cells, Csk is constitutively present in lipid rafts through an interaction with the Csk SH2-binding protein, PAG, also known as Cbp. Upon triggering of the T cell antigen receptor (TCR), PAG/Cbp is rapidly dephosphorylated leading to dissociation of Csk from lipid rafts. However, tyrosine phosphorylation of PAG/Cbp resumes after 3--5 min, at which time Csk reassociates with the rafts. Cells overexpressing a mutant Csk that lacks the catalytic domain, but displaces endogenous Csk from lipid rafts, have elevated basal levels of TCR-zeta-chain phosphorylation and spontaneous activation of an NFAT-AP1 reporter from the proximal interleukin-2 promoter as well as stronger and more sustained responses to TCR triggering than controls. We suggest that a transient release from Csk-mediated inhibition by displacement of Csk from lipid rafts is important for normal T cell activation.
Asunto(s)
Activación de Linfocitos/fisiología , Microdominios de Membrana/fisiología , Proteínas Tirosina Quinasas/metabolismo , Linfocitos T/inmunología , Antígenos Transformadores de Poliomavirus/genética , Proteína Tirosina Quinasa CSK , Células Cultivadas , Humanos , Células Jurkat , Modelos Biológicos , Muromonab-CD3/farmacología , Fosforilación , Fosfotirosina/metabolismo , Proteínas Tirosina Quinasas/genética , Receptores de Antígenos de Linfocitos T/fisiología , Proteínas Recombinantes/metabolismo , Linfocitos T/efectos de los fármacos , Transfección , Vanadatos/farmacología , Dominios Homologos src , Familia-src QuinasasRESUMEN
T cell receptor (TCR)-interacting molecule (TRIM) is a recently identified transmembrane adaptor protein, which is exclusively expressed in T cells. Here we demonstrate that in mature T cells, TRIM preferentially interacts with the TCR via the TCR-zeta chains and to a lesser extent via the CD3-straightepsilon/gamma heterodimer. Transient or stable overexpression of TRIM in Jurkat T cells results in enhancement of TCR expression on the cell surface and elevated induction of Ca(2+) mobilization after T cell activation. TRIM-mediated upregulation of TCR expression results from inhibition of spontaneous TCR internalization and stabilization of TCR complexes on the cell surface. Collectively, our data identify TRIM as a novel integral component of the TCR complex and suggest that one function of TRIM might be to modulate the strength of signals transduced through the TCR through regulation of TCR expression on the cell surface.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/fisiología , Proteínas de la Membrana/química , Proteínas de la Membrana/fisiología , Receptores de Antígenos de Linfocitos T/química , Receptores de Antígenos de Linfocitos T/fisiología , Transducción de Señal , Secuencia de Aminoácidos , Animales , Células COS , Calcio/metabolismo , Dimerización , Humanos , Células Jurkat , Datos de Secuencia MolecularRESUMEN
Mass spectrometry was applied to identify protein spots excised from an archived two-dimensional polyacrylamide gel that had been dried and stored for eight years at room temperature. All proteins were successfully identified. Detailed characterization of protein digests by matrix-assisted laser desorption/ionization (MALDI) peptide mapping, nanoelectrospray tandem mass spectrometry and MALDI-quadrupole time-of-flight mass spectrometry revealed no evidence of protein degradation or modifications that could hamper identification of proteins in a sequence database. The experiment with a model protein demonstrated that the pattern of tryptic peptides and the yield of individual peptides were not noticeably changed in the in-gel digest of the archived protein spot compared to the digest of the spot excised from a fresh gel. Thus, the characterization of "archived proteomes" has the potential to advance proteomic research without repeating "wet" biochemistry experiments, that had been perfected in the laboratory years ago.
Asunto(s)
Resinas Acrílicas , Proteoma/análisis , Secuencia de Aminoácidos , Humanos , Datos de Secuencia Molecular , Mapeo Peptídico/métodos , Procesamiento Proteico-Postraduccional , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Tripsina/metabolismoAsunto(s)
Proteínas Tirosina Fosfatasas/química , Proteínas Tirosina Fosfatasas/fisiología , Secuencia de Aminoácidos , Animales , Células Madre Hematopoyéticas/inmunología , Humanos , Ratones , Datos de Secuencia Molecular , Proteínas Tirosina Fosfatasas/genética , Proteínas Tirosina Fosfatasas Clase 3 Similares a Receptores , Homología de Secuencia de AminoácidoRESUMEN
The snake venom toxin convulxin activates platelets through the collagen receptor glycoprotein VI (GPVI)/Fc receptor gamma-chain (FcR gamma-chain) complex leading to tyrosine phosphorylation and activation of the tyrosine Syk and phospholipase Cgamma2 (PLCgamma2). In the present study, we demonstrate that convulxin is a considerably more powerful agonist than collagen or the GPVI-selective collagen-related peptide (CRP). Confirmation that the response to convulxin is mediated solely via Syk was provided by studies on Syk-deficient platelets. The increase in phosphorylation of the FcR gamma-chain is associated with marked increases in tyrosine phosphorylation of downstream proteins including Syk, linker for activation of T cells (LAT), SLP-76, and PLCgamma2. The transmembrane adapter LAT coprecipitates with SLP-76 and PLCgamma2, as well as with a number of other adapter proteins, some of which have not been previously described in platelets, including Cbl, Grb2, Gads, and SKAP-HOM. Gads is constitutively associated with SLP-76 and is probably the protein bridging its association with LAT. There was no detectable association between Grb2 and SLP-76 in control or stimulated cells, suggesting that the interaction of LAT with Grb2 is present in a separate complex to that of LAT-Gads-SLP-76. These results show that the trimeric convulxin stimulates a much greater phosphorylation of the FcR gamma-chain and subsequent downstream responses relative to CRP and collagen, presumably because of its ability to cause a greater degree of cross-linking of GPVI. The adapter LAT appears to play a critical role in recruiting a number of other adapter proteins to the surface membrane in response to activation of GPVI, presumably at sites of glycolipid-enriched microdomains, enabling an organized signaling cascade that leads to platelet activation.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Plaquetas/efectos de los fármacos , Proteínas Portadoras/fisiología , Venenos de Crotálidos/farmacología , Integrinas/metabolismo , Lectinas Tipo C , Proteínas de la Membrana , Fosfoproteínas/fisiología , Plaquetas/metabolismo , Colágeno/farmacología , Proteína Receptora de AMP Cíclico/farmacología , Proteína Adaptadora GRB2 , Humanos , Péptidos y Proteínas de Señalización Intracelular , Fosforilación , Proteínas/fisiología , Receptores de Colágeno , Receptores de IgG/fisiología , Fosfolipasas de Tipo C/fisiologíaRESUMEN
BACKGROUND: After allogeneic peripheral blood progenitor cell (PBPC) transplantation, a patient developed a severe hemolytic transfusion reaction due to passenger lymphocyte syndrome. CASE REPORT: A 50-year-old woman with secondary acute myeloid leukemia transforming from a myelodysplastic syndrome received an ABO-compatible PBPC graft from her HLA-identical sister. For prophylaxis of GVHD, the patient was treated with cyclosporine and methotrexate. Eighteen days after the transplant, the patient experienced a severe hemolytic transfusion reaction due to an alloantibody (anti-Jk(a)) produced by donor lymphocytes. RESULTS: The patient was typed as group A, Jk(a+) before transplantation; the donor was typed as group A, Jk(a-). On Day 18 after transplantation, the immunohematologic screening revealed a positive DAT (C3d 3+) and an alloanti-Jk(a). Hemolysis in the patient at that time was indicated by a drop in the Hb and an increase in the LDH level (maximum, 592 IU/L on Day 23). CONCLUSION: The course of hemolysis and the time of appearance of an alloantibody in this patient meet the criteria for passenger lymphocyte syndrome. In most cases, this syndrome is triggered by ABO system antibodies. This is the first reported case of passenger lymphocyte syndrome after PBPC transplantation that was due to an alloantibody that did not belong to the ABO system.
Asunto(s)
Anemia Hemolítica/etiología , Linfocitos B/inmunología , Incompatibilidad de Grupos Sanguíneos/etiología , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Isoanticuerpos/inmunología , Sistema del Grupo Sanguíneo de Kidd/inmunología , Enfermedad Aguda , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfocitos B/trasplante , Tipificación y Pruebas Cruzadas Sanguíneas , Separación Celular , Terapia Combinada , Transfusión de Eritrocitos/efectos adversos , Femenino , Reacción Injerto-Huésped , Humanos , Isoanticuerpos/biosíntesis , Leucemia Mieloide/terapia , Depleción Linfocítica , Persona de Mediana Edad , Síndromes Mielodisplásicos/terapia , Trasplante HomólogoRESUMEN
According to a recently proposed hypothesis, initiation of signal transduction via immunoreceptors depends on interactions of the engaged immunoreceptor with glycosphingolipid-enriched membrane microdomains (GEMs). In this study, we describe a novel GEM-associated transmembrane adaptor protein, termed phosphoprotein associated with GEMs (PAG). PAG comprises a short extracellular domain of 16 amino acids and a 397-amino acid cytoplasmic tail containing ten tyrosine residues that are likely phosphorylated by Src family kinases. In lymphoid cell lines and in resting peripheral blood alpha/beta T cells, PAG is expressed as a constitutively tyrosine-phosphorylated protein and binds the major negative regulator of Src kinases, the tyrosine kinase Csk. After activation of peripheral blood alpha/beta T cells, PAG becomes rapidly dephosphorylated and dissociates from Csk. Expression of PAG in COS cells results in recruitment of endogenous Csk, altered Src kinase activity, and impaired phosphorylation of Src-specific substrates. Moreover, overexpression of PAG in Jurkat cells downregulates T cell receptor-mediated activation of the transcription factor nuclear factor of activated T cells. These findings collectively suggest that in the absence of external stimuli, the PAG-Csk complex transmits negative regulatory signals and thus may help to keep resting T cells in a quiescent state.