Downstream Signaling Molecules Bind to Different Phosphorylated Immunoreceptor Tyrosine-based Activation Motif (ITAM) Peptides of the High Affinity IgE Receptor

Kimura, Takeshi; Kihara, H.; Bhattacharyya, Siba P.; Sakamoto, Hiroshi; Appella, Ettore; Siraganian, Reuben P.

doi:10.1074/jbc.271.44.27962

Cited by 68 publications

(67 citation statements)

References 61 publications

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“…Therefore, the recruitment of SHP-2 to the membrane by binding to the tyrosine-phosphorylated PECAM-1 could enhance its phosphatase activity by these two mechanisms. The recruitment of SHP-2 to PECAM-1 would bring it in close proximity to other potential substrates such as Fc⑀RI, the Src family tyrosine kinase Lyn, the proteintyrosine kinase Syk, the adaptor protein Shc, phospholipase C␥1, and of the SH2 domain-containing inositol polyphosphate 5-phosphatase, SHIP (26,50). Thus, SHP-2 could regulate the extent of the tyrosine phosphorylation of these molecules in receptor-activated cells thereby controlling the resulting downstream signals.…”

Section: Discussionmentioning

confidence: 99%

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The Protein-tyrosine Phosphatase SHP-2 Associates with Tyrosine-phosphorylated Adhesion Molecule PECAM-1 (CD31)

Sagawa

Kimura

Swieter

et al. 1997

Journal of Biological Chemistry

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101

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Aggregation of many cell-surface receptors results in tyrosine phosphorylation of numerous proteins. We previously observed the tyrosine phosphorylation of the platelet/endothelial cell adhesion molecule, PECAM-1 (CD31), after Fc⑀RI stimulation in rat basophilic leukemia RBL-2H3 cells. Here we found that PECAM-1 was also transiently tyrosine-phosphoryated after adherence of these cells to fibronectin. Similarly aggregation of the T cell receptor on Jurkat cells also induced this tyrosine phosphorylation. The protein-tyrosine phosphatase SHP-2 is a widely expressed cytosolic enzyme with two Src homology 2 (SH2) domains. SHP-2, but not the related protein-tyrosine phosphatase SHP-1, associated with PECAM-1. This association of the two proteins correlated with the extent of the tyrosine phosphorylation of PECAM-1. A fusion protein containing the two SH2 domains of SHP-2 precipitated PECAM-1 from cell lysates and also directly bound to phosphorylated PECAM-1. In immune precipitate phosphatase assays, there was tyrosine dephosphorylation of PECAM-1. Therefore, integrin and immune receptor activation results in tyrosine phosphorylation of PECAM-1 and the binding of the protein-tyrosine phosphatase SHP-2, which could regulate receptor-mediated signaling in cells.

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Section: Discussionmentioning

confidence: 99%

“…All other materials not indicated under "Experimental Procedures" were described previously (7,25,26).…”

Section: Methodsmentioning

confidence: 99%

The Protein-tyrosine Phosphatase SHP-2 Associates with Tyrosine-phosphorylated Adhesion Molecule PECAM-1 (CD31)

Sagawa

Kimura

Swieter

et al. 1997

Journal of Biological Chemistry

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101

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“…ITAM Binding Assay-Biotinylated peptides (phosphorylated or non-phosphorylated) corresponding to the ITAM of the ␥-subunit of Fc⑀RI were described previously (18). Cells lysates prepared with the Triton lysis buffer described above were precleared by mixing for 90 min at 4°C with streptavidinagarose beads.…”

Section: Syk Autophosphorylation and In Vitro Kinasementioning

confidence: 99%

Tyrosines in the Carboxyl Terminus Regulate Syk Kinase Activity and Function

Castro

Zhang

Jamur

et al. 2010

Journal of Biological Chemistry

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The Syk tyrosine kinase family plays an essential role in immunoreceptor tyrosine-based activation motif (ITAM) signaling. The binding of Syk to tyrosine-phosphorylated ITAM subunits of immunoreceptors, such as Fc⑀RI on mast cells, results in a conformational change, with an increase of enzymatic activity of Syk. This conformational change exposes the COOH-terminal tail of Syk, which has three conserved Tyr residues (Tyr-623, Tyr-624, and Tyr-625 of rat Syk). To understand the role of these residues in signaling, wild-type and mutant Syk with these three Tyr mutated to Phe was expressed in Syk-deficient mast cells. There was decreased Fc⑀RI-induced degranulation, nuclear factor for T cell activation and NFB activation with the mutated Syk together with reduced phosphorylation of MAP kinases p38 and p42/44 ERK. In non-stimulated cells, the mutated Syk was more tyrosine phosphorylated predominantly as a result of autophosphorylation. In vitro, there was reduced binding of mutated Syk to phosphorylated ITAM due to this increased phosphorylation. This mutated Syk from non-stimulated cells had significantly reduced kinase activity toward an exogenous substrate, whereas its autophosphorylation capacity was not affected. However, the kinase activity and the autophosphorylation capacity of this mutated Syk were dramatically decreased when the protein was dephosphorylated before the in vitro kinase reaction. Furthermore, mutation of these tyrosines in the COOH-terminal region of Syk transforms it to an enzyme, similar to its homolog ZAP-70, which depends on other tyrosine kinases for optimal activation. In testing Syk mutated singly at each one of the tyrosines, Tyr-624 but especially Tyr-625 had the major role in these reactions. Therefore, these results indicate that these tyrosines in the tail region play a critical role in regulating the kinase activity and function of Syk.Syk and ZAP-70 are members of a tyrosine kinase family one or both of which are expressed in most hematopoietic cells (1-5). Structurally Syk/ZAP-70 consists of two Src-homology 2 domains (SH2) 3 and a kinase domain followed by a short COOH-terminal extension (tail). The inter-SH2 domain is located between the NH 2 -terminal SH2 (SH2-N) and COOHterminal SH2 (SH2-C), whereas the linker region is between the SH2-C and the kinase domains (6, 7). An alternatively spliced form of Syk, termed SykB, lacks a 23-amino acid sequence in the linker domain (8). Although both isoforms are expressed in immune cells such as the RBL-2H3 rat mast cell line, Syk is more efficient than SykB in immunoreceptor signaling (8,9). Syk is expressed in B cells, mast cells, immature T cells, and platelets, whereas ZAP-70 is in T cells and natural killer cells (3, 10). Syk and ZAP-70 are essential for signal transduction from cell surface immunoreceptors (10 -12). These receptors on many hematopoietic cells such as T, B, and mast cells have subunits, which contain a sequence named the immunoreceptor tyrosine-based activation motif (ITAM) which contains two tyrosine residues (13...

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“…A mathematical analysis of data from the LynFc⑀RI transfectants indicated a single Lyn molecule was sufficient to phosphorylate a dimerized receptor (12). Yeast two-hybrid studies (8) indicated that the unique domain of both Lyn A and Lyn B could bind directly to the unphosphorylated C-terminal cytoplasmic domain of the Fc⑀RI ␤-chain, while peptide-binding studies (9,13) indicated that the SH2 domain of Lyn bound to phosphorylated Fc⑀RI ␤ ITAMs. Furthermore, chemical cross-linking studies demonstrated that 5% of total cellular Lyn in rat basophilic leukemia (RBL)-2H3 cells was bound to resting, unphosphorylated receptors (7).…”

Section: Regulation Of Rat Basophilic Leukemia-2h3 Mast Cellmentioning

confidence: 99%

“…We next examined the consequences of Lyn unique domain transfection on IgE receptor-mediated tyrosine phosphorylation of Syk kinase, which is recruited to phosphorylated Fc⑀RI ␥ ITAMs via its tandem SH2 domains after Fc⑀RI aggregation (9,13). Control and Lyn unique domain transfectants were sensitized as before with DNP-specific IgE and stimulated with 50 ng/ml DNP 12 -BSA Ag, and the Syk protein immunoprecipitated with polyclonal antiSyk.…”

Section: Kinetics Of Syk Tyrosine Phosphorylation Induced By Fc⑀ri Agmentioning

confidence: 99%

Regulation of Rat Basophilic Leukemia-2H3 Mast Cell Secretion by a Constitutive Lyn Kinase Interaction with the High Affinity IgE Receptor (FcεRI)

Vonakis

Gibbons

Rotté

et al. 2005

The Journal of Immunology

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Signaling through the high affinity IgE receptor is initiated by noncovalently associated Lyn kinase, resulting in the secretion of inflammatory mediators from mast cells. A fraction of the total cellular Lyn is associated via its N-terminal unique domain with the cytoplasmic domain of the FcεRI β subunit before receptor aggregation. In the current study, we stably transfected the unique domain of Lyn into rat basophilic leukemia-2H3 mast cells and examined the consequences on FcεRI-induced signal transduction and mediator secretion to further define the role of the unique domain of Lyn in mast cell secretion. Tyrosine phosphorylation of FcεRI β and γ subunits was partially inhibited in the Lyn unique domain transfectants after Ag stimulation. Ag stimulation of Lyn unique domain transfectants was accompanied by enhanced phosphorylation of MEK and ERK-2, which are required for leukotriene C4 (LTC4) release, and production of LTC4 was increased 3- to 5-fold, compared with cells transfected with vector alone. Conversely, tyrosine phosphorylation of the adaptor protein Gab2, which is essential for mast cell degranulation, was inhibited after Ag stimulation of Lyn unique domain transfectants, and Ag-induced release of histamine was inhibited up to 48%. In rat basophilic leukemia-2H3 cells, Lyn thus plays a dual role by positively regulating FcεRI phosphorylation and degranulation while negatively regulating LTC4 production. This study provides further evidence that the constitutive interaction between the unique domain of Lyn and the FcεRI β subunit is a crucial step in the initiation of FcεRI signaling and that Lyn is limiting for FcεRI-induced secretion of inflammatory mediators.

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Downstream Signaling Molecules Bind to Different Phosphorylated Immunoreceptor Tyrosine-based Activation Motif (ITAM) Peptides of the High Affinity IgE Receptor

Cited by 68 publications

References 61 publications

The Protein-tyrosine Phosphatase SHP-2 Associates with Tyrosine-phosphorylated Adhesion Molecule PECAM-1 (CD31)

The Protein-tyrosine Phosphatase SHP-2 Associates with Tyrosine-phosphorylated Adhesion Molecule PECAM-1 (CD31)

Tyrosines in the Carboxyl Terminus Regulate Syk Kinase Activity and Function

Regulation of Rat Basophilic Leukemia-2H3 Mast Cell Secretion by a Constitutive Lyn Kinase Interaction with the High Affinity IgE Receptor (FcεRI)

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