Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor

F. Yanaga, A. Poole, J. Asselin, R. Blake, G. L. Schieven, E. A. Clark, C. L. Law, S. P. Watson

研究成果: ジャーナルへの寄稿記事

123 引用 (Scopus)

抄録

Activation of human platelets by cross-linking of the platelet low-affinity IgG receptor, the Fcγ receptor IIA (Fcγ-RIIA), or by collagen is associated with rapid phosphorylation on tyrosine of the non-receptor tyrosine kinase syk. Phosphorylation is still observed, albeit sometimes reduced, in the presence of a combination of a protein kinase C inhibitor, Ro 31-8220, and the intracellular calcium chelator, BAPTA-AM, demonstrating independence from phosphoinositide-specific phospholipase C (PLC) activity. In contrast, the combination of Ro 31-8220 and BAPTA-AM completely inhibits phosphorylation of syk in thrombin-stimulated platelets. Phosphorylation of syk increases its autophosphorylation activity measured in a kinase assay performed on syk immunoprecipitates. Fcγ-RIIA also undergoes phosphorylation in syk immunoprecipitates from platelets activated by cross-linking of Fcγ-RIIA but not by collagen, suggesting that it associates with the kinase. Consistent with this, tyrosine-phosphorylated Fcγ-RIIA is precipitated by a glutathione S-transferase (GST) fusion protein containing the tandem src homology (SH2) domains of syk from Fcγ-RIIA-but not collagen-activated cells. Two uncharacterized tyrosine-phosphorylated proteins of 40 and 65 kDa are uniquely precipitated by a GST fusion protein containing the tandem syk-SH2 domains in collagen-stimulated platelets. A peptide based on the antigen recognition activation motif(ARAM) of Fcγ-RIIA, and phosphorylated on the two tyrosine residues found within this region, selectively binds syk from lysates of resting platelets; this interaction is not seen with a non-phosphorylated peptide. Kinase assays on Fcγ-RIIA immunoprecipitates reveal the constitutive association of an unidentified kinase activity in resting cells which phosphorylates a 67 kDa protein. Syk is not detected in Fcγ-RIIA immunoprecipitates from resting cells but associates with the receptor following activation and, together with Fcγ-RIIA, is phosphorylated in the kinase assay in vitro. These results demonstrate that syk is activated by Fcγ-RIIA cross-linking and collagen, independent of PLC, suggesting that it may have an important role in the early events associated with platelet activation. The association of syk with Fcγ-RIIA appears to be mediated through the tandem SH2 domains in syk and the ARAM motif of Fcγ-RIIA. A similar interaction may underlie the response to collagen, suggesting that its signalling receptor contains an ARAM motif.

元の言語英語
ページ(範囲)471-478
ページ数8
ジャーナルBiochemical Journal
311
発行部数2
DOI
出版物ステータス出版済み - 1 1 1995

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Fc Receptors
Platelets
Tyrosine
Collagen
Blood Platelets
Phosphorylation
Proteins
src Homology Domains
Chemical activation
Phosphotransferases
Assays
Platelet Activation
Type C Phospholipases
Glutathione Transferase
Antigens
Fusion reactions
Association reactions
Phosphoinositide Phospholipase C
IgG Receptors
Peptides

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Yanaga, F., Poole, A., Asselin, J., Blake, R., Schieven, G. L., Clark, E. A., ... Watson, S. P. (1995). Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor. Biochemical Journal, 311(2), 471-478. https://doi.org/10.1042/bj3110471

Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor. / Yanaga, F.; Poole, A.; Asselin, J.; Blake, R.; Schieven, G. L.; Clark, E. A.; Law, C. L.; Watson, S. P.

:: Biochemical Journal, 巻 311, 番号 2, 01.01.1995, p. 471-478.

研究成果: ジャーナルへの寄稿記事

Yanaga, F, Poole, A, Asselin, J, Blake, R, Schieven, GL, Clark, EA, Law, CL & Watson, SP 1995, 'Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor', Biochemical Journal, 巻. 311, 番号 2, pp. 471-478. https://doi.org/10.1042/bj3110471
Yanaga, F. ; Poole, A. ; Asselin, J. ; Blake, R. ; Schieven, G. L. ; Clark, E. A. ; Law, C. L. ; Watson, S. P. / Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor. :: Biochemical Journal. 1995 ; 巻 311, 番号 2. pp. 471-478.
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abstract = "Activation of human platelets by cross-linking of the platelet low-affinity IgG receptor, the Fcγ receptor IIA (Fcγ-RIIA), or by collagen is associated with rapid phosphorylation on tyrosine of the non-receptor tyrosine kinase syk. Phosphorylation is still observed, albeit sometimes reduced, in the presence of a combination of a protein kinase C inhibitor, Ro 31-8220, and the intracellular calcium chelator, BAPTA-AM, demonstrating independence from phosphoinositide-specific phospholipase C (PLC) activity. In contrast, the combination of Ro 31-8220 and BAPTA-AM completely inhibits phosphorylation of syk in thrombin-stimulated platelets. Phosphorylation of syk increases its autophosphorylation activity measured in a kinase assay performed on syk immunoprecipitates. Fcγ-RIIA also undergoes phosphorylation in syk immunoprecipitates from platelets activated by cross-linking of Fcγ-RIIA but not by collagen, suggesting that it associates with the kinase. Consistent with this, tyrosine-phosphorylated Fcγ-RIIA is precipitated by a glutathione S-transferase (GST) fusion protein containing the tandem src homology (SH2) domains of syk from Fcγ-RIIA-but not collagen-activated cells. Two uncharacterized tyrosine-phosphorylated proteins of 40 and 65 kDa are uniquely precipitated by a GST fusion protein containing the tandem syk-SH2 domains in collagen-stimulated platelets. A peptide based on the antigen recognition activation motif(ARAM) of Fcγ-RIIA, and phosphorylated on the two tyrosine residues found within this region, selectively binds syk from lysates of resting platelets; this interaction is not seen with a non-phosphorylated peptide. Kinase assays on Fcγ-RIIA immunoprecipitates reveal the constitutive association of an unidentified kinase activity in resting cells which phosphorylates a 67 kDa protein. Syk is not detected in Fcγ-RIIA immunoprecipitates from resting cells but associates with the receptor following activation and, together with Fcγ-RIIA, is phosphorylated in the kinase assay in vitro. These results demonstrate that syk is activated by Fcγ-RIIA cross-linking and collagen, independent of PLC, suggesting that it may have an important role in the early events associated with platelet activation. The association of syk with Fcγ-RIIA appears to be mediated through the tandem SH2 domains in syk and the ARAM motif of Fcγ-RIIA. A similar interaction may underlie the response to collagen, suggesting that its signalling receptor contains an ARAM motif.",
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T1 - Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fcγ-IIA receptor

AU - Yanaga, F.

AU - Poole, A.

AU - Asselin, J.

AU - Blake, R.

AU - Schieven, G. L.

AU - Clark, E. A.

AU - Law, C. L.

AU - Watson, S. P.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Activation of human platelets by cross-linking of the platelet low-affinity IgG receptor, the Fcγ receptor IIA (Fcγ-RIIA), or by collagen is associated with rapid phosphorylation on tyrosine of the non-receptor tyrosine kinase syk. Phosphorylation is still observed, albeit sometimes reduced, in the presence of a combination of a protein kinase C inhibitor, Ro 31-8220, and the intracellular calcium chelator, BAPTA-AM, demonstrating independence from phosphoinositide-specific phospholipase C (PLC) activity. In contrast, the combination of Ro 31-8220 and BAPTA-AM completely inhibits phosphorylation of syk in thrombin-stimulated platelets. Phosphorylation of syk increases its autophosphorylation activity measured in a kinase assay performed on syk immunoprecipitates. Fcγ-RIIA also undergoes phosphorylation in syk immunoprecipitates from platelets activated by cross-linking of Fcγ-RIIA but not by collagen, suggesting that it associates with the kinase. Consistent with this, tyrosine-phosphorylated Fcγ-RIIA is precipitated by a glutathione S-transferase (GST) fusion protein containing the tandem src homology (SH2) domains of syk from Fcγ-RIIA-but not collagen-activated cells. Two uncharacterized tyrosine-phosphorylated proteins of 40 and 65 kDa are uniquely precipitated by a GST fusion protein containing the tandem syk-SH2 domains in collagen-stimulated platelets. A peptide based on the antigen recognition activation motif(ARAM) of Fcγ-RIIA, and phosphorylated on the two tyrosine residues found within this region, selectively binds syk from lysates of resting platelets; this interaction is not seen with a non-phosphorylated peptide. Kinase assays on Fcγ-RIIA immunoprecipitates reveal the constitutive association of an unidentified kinase activity in resting cells which phosphorylates a 67 kDa protein. Syk is not detected in Fcγ-RIIA immunoprecipitates from resting cells but associates with the receptor following activation and, together with Fcγ-RIIA, is phosphorylated in the kinase assay in vitro. These results demonstrate that syk is activated by Fcγ-RIIA cross-linking and collagen, independent of PLC, suggesting that it may have an important role in the early events associated with platelet activation. The association of syk with Fcγ-RIIA appears to be mediated through the tandem SH2 domains in syk and the ARAM motif of Fcγ-RIIA. A similar interaction may underlie the response to collagen, suggesting that its signalling receptor contains an ARAM motif.

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