Physiological role of endogenous mono (ADP-ribosyl) ation of GTP binding proteins

GTP 结合蛋白内源性单 (ADP-核糖基) 化的生理作用

• 批准号: 02807205
• 负责人: TANUMA Sei-ichi
• 金额: $ 0.96万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02807205/
• 关键词: mono (ADP-ribosyl) ation; mono (ADP-ribosyl) transferase; G protein; adenylate cyclase; platelet; signal transduction; アデニレ-トシクラ-ゼ; モノ(ADPーリボ-ス)グリコヒドロラ-ゼ

Three types of arginine-specific ADP-ribosyltransferase, named ADP-ribosyltransferase A, were purified from human platelets using polyarginine as an ADP-ribose acceptor. When human platelet membranes were incubated with the transferase A in the presence of [32P]-NAD, a 45 K Membrane protein was specifically labeled. On the analysis of labeled product with phosphodiesterase by strong-anion exchange HPLC, more than 90% of the radioactivity was deleted as 5'-AMP. The linkage between mono (ADP-ribose) and 45K protein was sensitive to neutral hydroxylamine treatment. ADP-ribose transfer to 45K protein by this enzyme was suppressed when membranes were pre-ADP-ribosylated by chlera toxin. These results suggest that the ADP-ribosyltransferase AI catalyzes mono (ADP-ribosyl) ation of the specific membrane G protein, Gs.Incubation of platelet membranes with the transferase AI resulted in activation of the adenylate cyclase system. This stimulatory effects of the transferase AI on the adenylate cyclase system was further enhanced by prostagrandin (PG) El treatment.These results indicate a role of ADP-ribosyltransferase AI in regulation of the adentlate cyclase system via endogenous mono (ADP-ribosyl) ation og Gs.

以多聚精氨酸为ADP核糖受体，从人血小板中分离纯化了三种精氨酸特异性ADP核糖基转移酶，命名为ADP核糖基转移酶A。当人血小板膜与转移酶A在[32P]-NAD存在下孵育时，45K膜蛋白被特异性标记。对磷酸二酯酶标记产物进行强阴离子交换高效液相色谱分析时，90%以上的放射性被去除为5‘-AMP。ADP-核糖与45K蛋白的连接对中性羟胺敏感。当膜被霍乱毒素预先ADP-核糖化时，该酶能抑制ADP-核糖向45K蛋白的转移。这些结果表明，ADP-核糖基转移酶AI催化特异性膜G蛋白G的单(ADP-核糖基)反应。转移酶AI与血小板膜结合导致腺苷环化酶系统的激活。前列腺素(PG)EL可进一步增强转移酶AI对腺苷环化酶系统的这种刺激作用。这些结果表明，ADP-核糖基转移酶AI通过内源性ADP-核糖基转移酶对腺苷环化酶系统的调节作用。

项目成果

Sei-ichi TANUMA: "Characterization of two forms of poly(ADP-ribose)glycohydrolase in guinea pig liver." Biochemistry. 30. 5907-5912 (1991)

Sei-ichi TANUMA：“豚鼠肝脏中两种形式的聚（ADP-核糖）糖水解酶的表征。”

Sei-ichi TANUMA: "Mono(ADP-ribosyl)ation of Gs by eukaryotic arginine-specific ADP-ribosyl-transferase stimulates adenylate cyclasesystem." Biochem. Biophys.Res.Commun.176. 1014-1019 (1991)

Sei-ichi TANUMA：“真核精氨酸特异性 ADP-核糖基转移酶对 G 进行单（ADP-核糖基）化会刺激腺苷酸环化酶系统。”

Seiーichi TANUMA: "Antitumor,Antiviral and Immunopotentiating activities of pine cone extracts:Potential medicinal efficacy of natural and synthetic ligninーrelatedーrelated materials." Anticancer Res.11. 881-888 (1991)

Seiichi TANUMA：“松果提取物的抗肿瘤、抗病毒和免疫增强活性：天然和合成木质素相关材料的潜在药用功效。” 881-888（1991）。

Sei-ichi TANUMA: "Antitumor,Antivial and Immunopotentiating activites of pine cone extracts:Potential medicinalefficacy of natural and synthetic lignin-related materials." Anticancer Res.11. 881-888 (1991)

Sei-ichi TANUMA：“松果提取物的抗肿瘤、抗病毒和免疫增强活性：天然和合成木质素相关材料的潜在药用功效。”

S. Tanuma: "Eukaryotic mono (ADP-Ribosyl) transferase that ADP-ribosylates the a-subunit of GTP-binding regulatory protein Gi." J. Biol. Chem.263. 5485-5489 (1988)

S. Tanuma：“真核单（ADP-核糖基）转移酶，ADP-核糖基化 GTP 结合调节蛋白 Gi 的 a 亚基。”