Characterization of 85Kd protein tyrosine phosphorylated by erythropoietin stimulation

促红细胞生成素刺激磷酸化的 85Kd 蛋白酪氨酸的表征

• 批准号: 05670926
• 负责人: KOMATSU Norio
• 金额: $ 1.41万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670926/
• 关键词: Erythropoietin; Signal transduction; Tyrosine phosphorylation; Phospholipase C-_<gamma>1; エリスロポエチン受容体

Erythropoietin (Epo) is the major regulator of the proliferation and differentiation of erythroid precursors through interaction with its receptor (Epo-R). Although Epo-R lacks a tyrosine kinase consensus sequence within its intracellular domain, the addition of its ligand to Epo-responsive cells, UT-7/Epo, induces the rapid and transient tyrosine phosphorylation of 145,130,80-85 and 40 Kd cellular proteins. Tyrosine phosphorylation of these proteins occurred dose- and time-dependently. We showed that the tyrosine phosphorylated 145 Kd protein is identical to PLC-_<gamma>1. Tyrosine phosphorylation of this protein is detectable within 30 seconds and almost reaches the maximum at 1 minute. This can last up to 10 minutes and declines thereafter. Additionally, in Epo-stimulated cells, PLC-_<gamma>1 become physically associated with 80-85 and 40 Kd proteins which have been tyrosine phosphorylated in response to Epo. We tried to identified the 80-85 Kd protein and found that this protein is not identical to P13K,Epo-R,nor Raf-1. We are now trying to identify this protein.

促红细胞生成素 (Epo) 是通过与其受体 (Epo-R) 相互作用来调节红系前体细胞增殖和分化的主要调节剂。尽管 Epo-R 在其胞内结构域内缺乏酪氨酸激酶共有序列，但将其配体添加到 Epo 反应性细胞 UT-7/Epo 中，可诱导 145、130、80-85 和 40 Kd 细胞蛋白快速瞬时酪氨酸磷酸化。这些蛋白质的酪氨酸磷酸化发生剂量和时间依赖性。我们发现酪氨酸磷酸化的 145 Kd 蛋白与 PLC-_<gamma>1 相同。该蛋白的酪氨酸磷酸化可在 30 秒内检测到，并在 1 分钟时几乎达到最大值。这种现象可持续长达 10 分钟，之后逐渐减弱。另外，在Epo刺激的细胞中，PLC-_<γ>1变得与80-85和40Kd蛋白质物理关联，这些蛋白质已响应Epo而被酪氨酸磷酸化。我们尝试鉴定80-85 Kd蛋白，发现该蛋白与P13K、Epo-R和Raf-1不同。我们现在正试图鉴定这种蛋白质。

项目成果

Moroi,M.,et al: "Phorbol ester treatment of two megakaryoblastic cell lines,CMK and UT-7,inchiees specific protein composition changes with non-coincident time-courses" Biochimica et Biophysica Acta. 1177. 139-146 (1993)

Moroi，M.，等人：“佛波酯处理两种巨核细胞系（CMK 和 UT-7），导致特定蛋白质组成随时间进程不一致而变化”Biochimica et Biophysicala Acta。

Ren, H-Y,et al.: "Erythropoietin induces tyrosine phosphorylation and activation of phospholipapse C-_<gamma> 1 in a human erythropoietin-dependent cell line." J.Biol.Chem.269. 19633-19638 (1994)

Ren, H-Y 等人：“促红细胞生成素在人促红细胞生成素依赖性细胞系中诱导酪氨酸磷酸化和磷脂酶 C-_<gamma> 1 的激活。”

Ren,H-Y.,et al: "Erythropoietin indnees tyrosine phosphorylation and activation of phospholipase C-γ1 in a human erythropoietin-dependent cell line" J.Biol.Chem.269. 19633-19638 (1994)

Ren, H-Y., 等人：“人促红细胞生成素依赖性细胞系中促红细胞生成素酪氨酸磷酸化和磷脂酶 C-γ1 的激活”J.Biol.Chem.269 (1994)。

Norio Komatsu: "Establishment and characterization of an erythropoietin-dependent subline,UT-7/Epo,derived from human leukemia cell line,UT-7" Blood. 82. 456-464 (1993)

Norio Komatsu：“源自人白血病细胞系 UT-7 的促红细胞生成素依赖性亚系 UT-7/Epo 的建立和表征”血液。

Komatsu,N.,et al: "Establishment and characterization of an crythropoietin-dependent subline,UT-7/Epo,derivedfrom human leukemia cell line.UT-7" Blood. 82. 456-464 (1993)

Komatsu, N., 等人：“源自人白血病细胞系的促红细胞生成素依赖性亚系 UT-7/Epo 的建立和表征。UT-7”血液。