1994 Fiscal Year Annual Research Report

免疫担当細胞における新しい非受容体型蛋白質チロシン燐酸化酵素P72^<syk>の作用

Research Project

Project/Area Number	05670117
Research Institution	Fukui Medical School
Principal Investigator	柳茂福井医科大学, 医学部, 助手 (60252003)
Co-Investigator(Kenkyū-buntansha)	山村博平福井医科大学, 医学部, 教授 (90030882)
Keywords	Syk / チロシンキナーゼ / 抗Syk抗体 / インテグリン / 免疫系細胞(B細胞) / B細胞抗原レセプター / Lyn / ジーンターゲッティング
Research Abstract	Porcine Sykの部分的な配列に対する抗体を複数作製することに成功した。さらにヒト及びマウスにおいてもSykを認識する抗体を作製することが出来た。これらを用いて当初の目的である細胞接着分子であるインテグリンとSykとの関係について血小板を用いて解析を進めた。その結果、Sykがインテグリンからのシグナルにおいて活性化し、さらにこれらと複合体を形成することが明らかとなり、細胞接着現象に深い関係にあることを示唆することが出来た。また黒崎らと我々のグループは免疫系細胞におけるSykの機能を調べるためにB細胞を用いてチキンのB細胞由来の培養細胞にジーンターゲッティングの手法を用いてSyk欠損B細胞およびLyn欠損B細胞を作製し、その機能的な差異を調べた。その結果、B細胞抗原レセプターからの情報伝達にSykとLynの両方が必要でかつSykとLynに機能的な差異を明らかにした。またIgM刺激依存性の細胞内チロシンリン酸化の亢進においてSykとSrcファミリーチロシンキナーゼとの間に相乗的効果が存在しており機能的相互作用を示唆することが出来た。以上の結果より、SykはB細胞抗原レセプターからの情報伝達において中心的な役割をしていることが明らかとなった。また以前より我々は血小板においてトロンビン刺激によりSykがわずか数秒で劇的に活性化することを見いだしたが、今回トロンビンだけでなくコラーゲン、PAF(Platelet activating factor)、Thromboxane A_2、WGA(Wheat germ aggulutinin)、Ca^<2+>イオンフォアなどもSykを活性化することを報告し、この酵素の普遍的な重要性を示唆した。さらにSykの細胞内分布に注目し解析を進めた結果、Sykは刺激依存性により細胞質から速やかに細胞膜や細胞骨格系分画に移行することを見いだし、血小板活性化におけるSykの関与を強く示唆した。

Research Products
(21 results)

All Other

All Publications (21 results)

[Publications] Minami,Y.: "Protein Tyrosine Kinase Syk Is Associated with and Activated by The IL‐2 Receptor;Possible Link with The c‐myc Induction Pathway.(in press)" Immunity.
[Publications] Nagai,K.: "Tyrosine phosphorylation of Shc is mediated through Lyn and Syk in B cell receptor signaing.(in press)" J.Biol.Chem.
[Publications] Qin,S.: "Activation and tyrosine phosphorylation of p72^<syk> as well as calcium mobilization after hydrogen peroxide stimulation in peripheral blood lymphocytes (in press)" Biochem.J.
[Publications] Takata,M.: "Tyrosine kinase Lyn and Syk regulate the B cell receptor‐coupled Ca^<2+> mobilization through the distinct pathway" EMBO J.13. 1341-1349 (1994)
[Publications] Sada,K.: "Activation of p72syk by thrombin in a cell‐free system." Biochem.Biophys.Res.Commun.200. 1-7 (1994)
[Publications] Yang,C.: "Purification and characterization of a protein‐tyrosine kinase p72syk from porcine spleen" Eur.J.Biochem.221. 973-978 (1994)
[Publications] Kurosaki,T.: "Syk activation by the src‐family tyrosine kinase in the B cell receptor signaling" J.Exp.Med.179. 1725-1729 (1994)
[Publications] Tsubokawa,M.: "CPTK71,a cytosolic protein‐tyrosine kinase previously purified from bovine platelets,is identical with p72syk." Biochem.Biophys.Res.Commun.200. 1592-1597 (1994)
[Publications] Qin,S.: "Interleukin 2 Mediates p72syk activation in peripheral blood lymphocytes." FEBS Lett.345. 233-236 (1994)
[Publications] Yanagi,S.: "Translocation,Activation,and Association with P13 Kinase of p72syk.are early Events during Platelet Activation." Eur.J.Biochem.224. 329-333 (1994)
[Publications] Fujii,C.: "Protein‐tyrosine kinase p72syk.is activated by collagen in platelets." Eur.J.Biochem.226. 243-248 (1994)
[Publications] Wang,X.: "Intracellular Calcium Dependent Activation of p72syk in Platelets" J.Biochem.116. 858-861 (1994)
[Publications] Nagai,K.: "p72syk is activated by vanadate plus H202 in porcine platelets and phosphorylates GTPase activating protein on tyrosine residue(s)" J.Biochem.116. 1176-1181 (1994)
[Publications] Tohyama,Y.: "Translocation of p72syk to the Cytoskeleton in Thrombin‐Stimulated Platelets." J.Biol.Chem.269. 32796-32799 (1994)
[Publications] Rezaul,K.: "Protein‐tyrosine kinase p72syk is activatedby platelet activating factor in platelets." Thrombosia and Haemostasis.72. 937-941 (1994)
[Publications] Minami,Y.: "Protein Tyrosine Kinase Syk Is Associated with and Activated by The IL‐2 Receptor;Possible Link with The c‐myc Induction Pathway.(in press)" Immunity.
[Publications] Nagai,K.: "Tyrosine phosphorylation of Shc is mediated through Lyn and Syk in B cell receptor signaing.(in press)" J.Biol.Chem.
[Publications] Qin,S.: "Activation and tyrosine phosphory lation of p72^<syk> as well as calcium mobilization after hydrogen peroxide stimulation in peripheral blood lymphocytes (in press)" Biochem.J.
[Publications] 稲津哲也: "実験医学" 羊土社, 2 (1994)
[Publications] 柳茂: "生化学" 日本生化学会, 11 (1994)
[Publications] Yamamura,H.: "The protein kinase Factsbook" Academic Press Limited (in press),

1994 Fiscal Year Annual Research Report

免疫担当細胞における新しい非受容体型蛋白質チロシン燐酸化酵素P72^<syk>の作用

Principal Investigator

柳 茂 福井医科大学, 医学部, 助手 (60252003)

Research Products

[Publications] Minami,Y.: "Protein Tyrosine Kinase Syk Is Associated with and Activated by The IL‐2 Receptor;Possible Link with The c‐myc Induction Pathway.(in press)" Immunity.

[Publications] Nagai,K.: "Tyrosine phosphorylation of Shc is mediated through Lyn and Syk in B cell receptor signaing.(in press)" J.Biol.Chem.

[Publications] Qin,S.: "Activation and tyrosine phosphorylation of p72^<syk> as well as calcium mobilization after hydrogen peroxide stimulation in peripheral blood lymphocytes (in press)" Biochem.J.

[Publications] Takata,M.: "Tyrosine kinase Lyn and Syk regulate the B cell receptor‐coupled Ca^<2+> mobilization through the distinct pathway" EMBO J.13. 1341-1349 (1994)

[Publications] Sada,K.: "Activation of p72syk by thrombin in a cell‐free system." Biochem.Biophys.Res.Commun.200. 1-7 (1994)

[Publications] Yang,C.: "Purification and characterization of a protein‐tyrosine kinase p72syk from porcine spleen" Eur.J.Biochem.221. 973-978 (1994)

[Publications] Kurosaki,T.: "Syk activation by the src‐family tyrosine kinase in the B cell receptor signaling" J.Exp.Med.179. 1725-1729 (1994)

[Publications] Tsubokawa,M.: "CPTK71,a cytosolic protein‐tyrosine kinase previously purified from bovine platelets,is identical with p72syk." Biochem.Biophys.Res.Commun.200. 1592-1597 (1994)

[Publications] Qin,S.: "Interleukin 2 Mediates p72syk activation in peripheral blood lymphocytes." FEBS Lett.345. 233-236 (1994)

[Publications] Yanagi,S.: "Translocation,Activation,and Association with P13 Kinase of p72syk.are early Events during Platelet Activation." Eur.J.Biochem.224. 329-333 (1994)

[Publications] Fujii,C.: "Protein‐tyrosine kinase p72syk.is activated by collagen in platelets." Eur.J.Biochem.226. 243-248 (1994)

[Publications] Wang,X.: "Intracellular Calcium Dependent Activation of p72syk in Platelets" J.Biochem.116. 858-861 (1994)

[Publications] Nagai,K.: "p72syk is activated by vanadate plus H202 in porcine platelets and phosphorylates GTPase activating protein on tyrosine residue(s)" J.Biochem.116. 1176-1181 (1994)

[Publications] Tohyama,Y.: "Translocation of p72syk to the Cytoskeleton in Thrombin‐Stimulated Platelets." J.Biol.Chem.269. 32796-32799 (1994)

[Publications] Rezaul,K.: "Protein‐tyrosine kinase p72syk is activatedby platelet activating factor in platelets." Thrombosia and Haemostasis.72. 937-941 (1994)

[Publications] Minami,Y.: "Protein Tyrosine Kinase Syk Is Associated with and Activated by The IL‐2 Receptor;Possible Link with The c‐myc Induction Pathway.(in press)" Immunity.

[Publications] Nagai,K.: "Tyrosine phosphorylation of Shc is mediated through Lyn and Syk in B cell receptor signaing.(in press)" J.Biol.Chem.

[Publications] Qin,S.: "Activation and tyrosine phosphory lation of p72^<syk> as well as calcium mobilization after hydrogen peroxide stimulation in peripheral blood lymphocytes (in press)" Biochem.J.

[Publications] 稲津 哲也: "実験医学" 羊土社, 2 (1994)

[Publications] 柳 茂: "生化学" 日本生化学会, 11 (1994)

[Publications] Yamamura,H.: "The protein kinase Factsbook" Academic Press Limited (in press),

柳茂福井医科大学, 医学部, 助手 (60252003)

[Publications] 稲津哲也: "実験医学" 羊土社, 2 (1994)

[Publications] 柳茂: "生化学" 日本生化学会, 11 (1994)