Effectcts of Ubiquitinated CD-10 and CD13 complex on HeLa, Raji and HL60.

泛素化 CD-10 和 CD13 复合物对 HeLa、Raji 和 HL60 的影响。

• 批准号: 11670421
• 负责人: ABE Sumiko
• 金额: $ 1.02万
• 依托单位: Fukushima Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670421/
• 关键词: Ubiquitin; CD10; CD13; Tyrosine-phosphorylation; semen; 精液

Human semen-specific γ-glutamyl transpeptidase active protein (SEGAP) is an enzyme complex of ubiquitinated CD10 and CD13. In this study, we investigated the potential of SEGAP to induce tyrosine-phosrchorylation in HeLa (human cervical carcinoma), Raji (human B cell lymphoma) and HL60 (human promyelocytic leukemia). FACS analysis of SEGAP-treated cells revealed that SEGAP may bind to these cells. Western blots of SEGAP-and anti-CD13 antibody-treated cell lysates were performed using an anti-phosphotyrosine antibody. Phosphorylation of 120-kDa and 55-kDa proteins in HeLa and HL60 were observed in 1-min and 5-min incubations with an anti-CD13 antibody. Since CD13 is not distributed in Raji, 120-kDa and 55-kDa proteins were not phosphorylated. SEGAP phosphorylated 110-kDa proteins during 1-min incubation in HeLa and 110-kDa, 70-kDa and 60-kDa proteins during 1-min incubation in HL60. These results suggested that SEGAP induces tyrosine-phosphorylations of 110-kDa, 70-kDa and 60-kDa proteins which are thought to be as PI3 kinase, Syk/ZAP-70 and Src family proteins, respectively. Messenger RNA levels of cytokines, lectins and apotosis related-proteins were also examined in SEGAP-treated HL60. SEGAP did not affect mRNA levels of IL1-β, IL8, Lyn, Fas and TNF.INF-γ and mannose-binding lectin RNAs were not expressed in HL60, and their levels were not affected by SEGAP.

人精液特异性γ-谷氨酰转肽酶活性蛋白是一种泛素化的CD10和CD13的酶复合体。在这项研究中，我们研究了SEGAP在HeLa(人宫颈癌)、Raji(人B细胞淋巴瘤)和HL60(人早幼粒细胞白血病)中诱导酪氨酸磷酸化的可能性。经SEGAP处理的细胞的FACS分析显示，SEGAP可能与这些细胞结合。用抗磷酸酪氨酸抗体对SEGAP和抗CD13抗体处理的细胞裂解产物进行Western blotting。抗CD13抗体作用1分钟和5分钟后，HeLa和HL60细胞中的120 kDa和55 kDa蛋白发生了磷酸化。由于CD13在Raji中没有分布，因此120 kDa和55 kDa的蛋白没有被磷酸化。SEGAP在HeLa中孵育1分钟可使110 kDa蛋白质磷酸化，在HL60中孵育1分钟可使110 kDa、70 kDa和60 kDa蛋白质磷酸化。这些结果表明，SEGAP诱导110 kDa、70 kDa和60 kDa蛋白的酪氨酸磷酸化，这些蛋白分别被认为是PI3激酶、Syk/ZAP-70和Src家族蛋白。SEGAP处理的HL60细胞中细胞因子、凝集素和细胞凋亡相关蛋白的信使RNA水平也被检测。SEGAP不影响IL-1-β、IL-8、LYN、Fas和TNFmRNA的表达，不表达INF-γ和甘露糖结合凝集素，其表达水平不受SEGAP的影响。

项目成果

Sumiko Abe, Miwako Suto, Kouichi Hiraiwa: "The effect of seminal γ-glutamyl transpeptidase active protein on tyrosine phosphorylation."Japanese Journal of Legal Medicine. 55(in print).

Sumiko Abe、Miwako Suto、Kouichi Hiraiwa：“精液 γ-谷氨酰转肽酶活性蛋白对酪氨酸磷酸化的影响。”《日本法律医学杂志》55（印刷中）。

阿部 すみ子: "癌化による精液特異γ-glutamyl transpeptidase構成ユビキチンの免疫反応性の変化"日本法医学雑誌. (印刷中).

Sumiko Abe：“由于癌变导致精液特异性 γ-谷氨酰转肽酶成分泛素的免疫反应性变化”，《日本法医学杂志》（正在出版）。

Abe Sumiko et al.: "Semen specific γ-glutamyl transpeptidase carries ABH antigen : a sandwich ELISA for simultaneous semen detection and its ABO blood typing"Clinica Chimica Acta. 283. 183-195 (1999)

Abe Sumiko 等：“精液特异性 γ-谷氨酰转肽酶携带 ABH 抗原：用于同步精液检测及其 ABO 血型的夹心 ELISA”Clinica Chimica Acta. 283. 183-195 (1999)

阿部すみ子 他: "精液特異γ-glutamyl transpeptidase活性酵素による培養細胞チロシン残基リン酸化"日本法医学雑誌. (印刷中).

Sumiko Abe 等人：“精液特异性 γ-谷氨酰转肽酶活性酶对培养细胞中酪氨酸残基的磷酸化”，《日本法医杂志》（出版中）。

Sumiko Abe, Hirobumi Gunji, Osamu Muraki Mikio Kuraya, Teizo Fujita, Miwako Suto Kouichi Hiraiwa: "Immunohistochemical characterization of seminal γ-glutamyl transpeptidase active protein consistuent-ubiquitin in prostate disease."Japanese Journal of Lega

Sumiko Abe、Hirobumi Gunji、Osamu Muraki Mikio Kuraya、Teizo Fujita、Miwako Suto Kouichi Hiraiwa：“前列腺疾病中精液 γ-谷氨酰转肽酶活性蛋白一致性泛素的免疫组织化学特征。”Japan Journal of Lega