Characterization of a new apoptosis-inducing protein from the cabbage butterfly

菜粉蝶中一种新的细胞凋亡诱导蛋白的表征

• 批准号: 13660344
• 负责人: WATANABE Masahiko
• 金额: $ 2.24万
• 依托单位: National Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660344/
• 关键词: cabbage butterfly; pierisin; ADP-rivosylation; DNA adduct; bacterial toxin; ADP-リボシル化

Pierisin-1 is a 98-kDa cytotoxic protein found in the cabbage butterfly and shows sequence similarity with ADP-ribosylating toxins such as cholera toxin. To identify a substrate protein for ADP-ribosyltransferase activity of pierisin-1, a mixture of[adenylate-^<32>P]-NAD, HeLa cell extract and pierisin-1 was incubated and analyzed by SDS/PAGE and autoradiography. The labeled material was protease resistant but DNase sensitive. This result suggested that DNA might be the substrate for ADP-ribosylation by pierisin. When a variety of oligonucleotides having different sequences were used for acceptors from the efficiency strongly correlated with the proportion of guanine base in each oligonucleotide. For structural determination of the reaction product by pierisin-1, 2'-deoxyguanosine was used as an acceptor molecule. The UV spectrum and ESI-MS suggested the presence of ADP-ribosylated dG. Several NMR analyses revealed the structure as α and β forms of N^2-(ADP-ribos-1-yl)-2'-deoxyguanosine. Finally, we independently synthesized chemically and confirmed the structural identity with the enzyme reaction product. When high-molecular-weight dsDNA was used as acceptor molecule, the same position of guanine base was ADP-ribosylated, as confirmed by structural analyses including ^1H-MNR. The ^<32>P-postlaveling method for detection of ADP-ribosylated dG indicated that ADP-ribosylation of N-2 of guanine base also occurred in HeLa cells treated with pierisin-1. From the above results, it is concluded that pierisin-1 transfers an ADP-ribosyl moiety from NAD to N-2 of guanine base of cellular DNA, and this causes apoptosis induction of the cells.

Pierisin-1是在卷心菜对接序列中发现的98 kDa细胞毒性蛋白，SITH ADP-核糖基毒素suckola sisola。 ]非SD/PAGE和PIERASIN-1的HELA细胞提取物和Pierisin-1诱导，但DNA敏感。用于来自每个Olighton核苷酸中的鸟嘌呤碱的疗效，用于质子素-1的结构。提出的是aDP-核糖基的DG。 ，鸟嘌呤碱的位置是ADP-核糖基化的MNR。细胞DNA的NAD​​至N-2，这会导致细胞中的Apoptosississ。

项目成果

Takamura-Enya, T.: "Mono(ADP-ribosyl)ation of 2'-deoxyguanosine residue in DNA by an apoptosis-inducing protein, pierisin-1, from cabbage butterfly"Proc.Natl.Acad.Sci.USA. 98. 12414-12419 (2001)

Takamura-Enya, T.：“来自菜蝴蝶的细胞凋亡诱导蛋白 Pierisin-1 对 DNA 中 2-脱氧鸟苷残基进行单（ADP-核糖基）化”Proc.Natl.Acad.Sci.USA。

Takeji Takamura Enya: "Mono(ADP-ribosyl)ation of 2'-deoxyguanosine residue in DNA by an apoptosis-inducing protein, pierisin-1, from cabbage butterfly."Proc Natl Acad Sci USA. 98. 12414-12419 (2001)

Takeji Takamura Enya：“来自菜蝴蝶的细胞凋亡诱导蛋白 Pierisin-1 对 DNA 中 2-脱氧鸟苷残基进行单（ADP-核糖基）化。”Proc Natl Acad Sci USA。

Kanazawa, T.: "Pierisin-1 from cabbage butterfly suppresses HeLa tumor growth in nude mice"Proc.Jpn.Acad.. 78B. 305-308 (2002)

Kanazawa, T.：“来自菜粉蝶的 Pierisin-1 抑制裸鼠 HeLa 肿瘤生长”Proc.Jpn.Acad.. 78B。

Takamura-Enya, T.: "Mono(ADP-ribosyl)ation of 2'-deoxyguanosine residue in DNA by an apoptosis-inducing protein, pierisin-1, from cabbage butterfly"Proc. Natl. Acad. Sci. USA. 98. 12414-12419 (2001)

Takamura-Enya, T.：“来自菜粉蝶的细胞凋亡诱导蛋白 Pierisin-1 对 DNA 中 2-脱氧鸟苷残基进行单（ADP-核糖基）化”Proc。

Matsushima-Hibiya, Y.: "Identification of glycosphingolipid receptors for pierisin-1, a guanine-specific ADP-ribosylating toxin from the cabbage butterfly"J. Biol. Chem.. (in press).

Matsushima-Hibiya, Y.：“Pierisin-1 的糖鞘脂受体的鉴定，Pierisin-1 是一种来自菜粉蝶的鸟嘌呤特异性 ADP 核糖基化毒素”J.