Reconstitution of High Affinity IgE Receptor-mediated Secretion By Transfecting Protein Tyrosine Kinase pp125FAK

Hamawy, Majed M.; Swieter, Mark; Mergenhagen, Stephan E.; Siraganian, Reuben P.

doi:10.1074/jbc.272.48.30498

Cited by 36 publications

(57 citation statements)

References 52 publications

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“…The cells were then incubated with NMG, anti-CD28 mAb, or anti-CD3 mAb, and the tubes were placed in a 37°C water bath for the indicated time. Following incubation, the cells were solubilized by adding an equal volume of 2ϫ ice-cold solubilizing buffer (10 mM Tris-HCl, pH 7.5, 1% Triton X-100, 0.5% sodium deoxycholate, 50 mM NaCl, 50 mM NaF, 2 mM Na 3 VO 4 , 0.5 unit/ml aprotinin, and 1 mM phenylmethylsulfonyl fluoride; final concentrations) and immediately placed on ice (65). The cells in the tubes were vortexed vigorously every 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…Although in adherent cells Fak is localized to focal adhesion sites, Pyk2 is mainly diffused throughout the cyto-plasm (49,50). Fak and Pyk2 become tyrosine-phosphorylated and activated after the stimulation of various receptors including TCR (44,45,47,(51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64), and both kinases have been linked to the signaling pathways that regulate MAP kinases (45,48,58,61,59).…”

mentioning

confidence: 99%

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CD28 Ligation Induces Tyrosine Phosphorylation of Pyk2 but Not Fak in Jurkat T Cells

Tsuchida

Knechtle

Hamawy

1999

Journal of Biological Chemistry

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Protein tyrosine kinases are critical for the function of CD28 in T cells. We examined whether the tyrosine kinases Pyk2 and Fak (members of the focal adhesion kinase family) are involved in CD28 signaling. We found that ligating CD28 in Jurkat T cells rapidly increases the tyrosine phosphorylation of Pyk2 but not of Fak. Paxillin, a substrate for Pyk2 and Fak, was not tyrosinephosphorylated after CD28 ligation. CD28-induced tyrosine phosphorylation of Pyk2 was markedly reduced in the absence of external Ca 2؉ . Previous studies have shown that the T cell antigen receptor (TCR) induces tyrosine phosphorylation of Pyk2. In this report, the concurrent ligation of CD28 and TCR increased tyrosine phosphorylation of Pyk2; however, the extent of phosphorylation by both receptors was equivalent to the sum of that induced by each receptor alone. The Syk/Zap inhibitor piceatannol blocked CD28, and TCR induced tyrosine phosphorylation of Pyk2, suggesting that Syk/ Zap is involved in Pyk2 phosphorylation. In contrast, the phosphatidylinositol 3-kinase inhibitor wortmannin blocked TCR-but not CD28-induced phosphorylation of Pyk2, suggesting that CD28 and TCR activate distinct pathways to induce tyrosine phosphorylation of Pyk2. Notably, depleting phorbol 12-myristate 13-acetate-sensitive protein kinase C did not block CD28-and CD3-induced tyrosine phosphorylation of Pyk2. These data provide evidence for the involvement of Pyk2 in the CD28 signaling cascade and suggest that neither Fak nor paxillin is involved in the signaling pathways of CD28.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

CD28 Ligation Induces Tyrosine Phosphorylation of Pyk2 but Not Fak in Jurkat T Cells

Tsuchida

Knechtle

Hamawy

1999

Journal of Biological Chemistry

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“…The stable transfection of FAK markedly enhances the Fc⑀RI-mediated secretion; this does not require the catalytic activity of the kinase, suggesting that FAK is functioning as an adaptor or linker molecule (22,35). This variant of the RBL-2H3 cells expressing low levels of FAK has a phenotype similar to some of the cultured EDMC that had decreased Fc⑀RI-mediated degranulation and could reflect cells that are transformed without some of the compensatory pathways present in the normally responsive cells.…”

Section: Signal Transduction In Fak-deficient Mast Cellsmentioning

confidence: 91%

“…Similarly, the stimulation of mast cells by Fc⑀RI aggregation results in the activation of these same pathways (34). There is also evidence from the rat basophilic leukemia RBL-2H3 cells of a role for FAK during the late stages of Fc⑀RI-induced degranulation (21,22,35). FAK may also link cell surface receptors to MAPK activation.…”

Section: Fc⑀ri Signaling In Fak-deficient Edmcmentioning

confidence: 99%

“…In previous experiments, a variant of the rat basophilic leukemia cell line was identified expressing low levels of FAK that had decreased Fc⑀RI, but not ionophore-mediated secretion (35). The stable transfection of FAK markedly enhances the Fc⑀RI-mediated secretion; this does not require the catalytic activity of the kinase, suggesting that FAK is functioning as an adaptor or linker molecule (22,35).…”

Section: Signal Transduction In Fak-deficient Mast Cellsmentioning

confidence: 99%

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Alterations in Granule Matrix and Cell Surface of Focal Adhesion Kinase-Deficient Mast Cells

Vial

Oliver

Jamur

et al. 2003

The Journal of Immunology

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Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine kinase that plays an important role in many cellular processes and is tyrosine phosphorylated after FcεRI aggregation in mast cells. In mice, null mutation of the fak gene results in a lethal phenotype in which the embryos fail to develop past day 8.5 of gestation. To study the role of FAK in these mast cells, 8.5-day embryos were isolated and placed in culture with IL-3 and stem cell factor (SCF). Although FAK was not required for the development of mast cells in culture, the FAK−/− embryo-derived mast cells had several distinct characteristics. Compared with the controls, the mast cells that lack FAK were less metachromatic and by electron microscopy had granules that appeared largely electron lucid, although their histamine content was unchanged. The FAK-deficient mast cells had a reduction in the content of chondroitin/dermatan sulfate, the major glycosaminoglycan component of the granular matrix. The FAK-deficient cells had fewer microvilli that were fused with each other, giving the cell surface a ruffled appearance. There was also a 3-fold increase in the number of cells highly expressing β7 integrin. However, signal transduction from the high affinity IgE receptor for the secretion of histamine was similar in the wild-type, heterozygote, and the FAK-deficient cells. The FcεRI-induced tyrosine phosphorylation of paxillin, Crk-associated tyrosine kinase substrate (CAS), and mitogen-activated protein kinase proteins was independent of FAK. These results indicate that FAK plays a role in regulating the glycosaminoglycan content of the secretory granules and influences the cell surface morphology of mast cells.

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Cyclical mechanical stretch enhances degranulation and IL‐4 secretion in RBL‐2H3 mast cells

Komiyama

Miyake

Asai

et al. 2013

Cell Biochemistry & Function

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Mast cells are widely distributed in the body and affect their surrounding environment through degranulation and secretion of cytokines. Conversely, mast cells are influenced by environmental stimuli such as cyclical mechanical stretch (CMS), such as that induced by heartbeat and respiration. Peripherally distributed mast cells are surrounded by extracellular matrix, where they bind IgE on their surface by expressing the high-affinity Fc receptor for IgE (FcεRI), and they release mediators after cross-linking of surface-bound IgE by allergen. To analyse how CMS affects mast cell responses, we examined the effect of applying CMS on the behaviour of IgE-bound mast cells (RBL-2H3 cell line) adhering to fibronectin as a substitute for extracellular matrix. We found that CMS enhanced FcεRI-mediated secretion in the presence of antigen (2,4-dinitrophenol-bovine serum albumin). CMS increased expression of IL-4 mRNA and secretion of IL-4 protein. Western blot analysis showed that CMS changes the signal transduction in mitogen-activated protein kinases and AKT, which in turn alters the regulation of IL-4 and increases the secretion of IL-4. These results suggest that CMS modulates the effect of mast cells on inflammation and resultant tissue remodelling. Understanding how CMS affects mast cell responses is crucial for developing therapies to treat mast cell-related diseases.

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Reconstitution of High Affinity IgE Receptor-mediated Secretion By Transfecting Protein Tyrosine Kinase pp125FAK

Cited by 36 publications

References 52 publications

CD28 Ligation Induces Tyrosine Phosphorylation of Pyk2 but Not Fak in Jurkat T Cells

CD28 Ligation Induces Tyrosine Phosphorylation of Pyk2 but Not Fak in Jurkat T Cells

Alterations in Granule Matrix and Cell Surface of Focal Adhesion Kinase-Deficient Mast Cells

Cyclical mechanical stretch enhances degranulation and IL‐4 secretion in RBL‐2H3 mast cells

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