Introduction of exogenous proteins into nucleus by using anthrax toxin

利用炭疽毒素将外源蛋白引入细胞核

• 批准号: 09557025
• 负责人: MEKADA Eisuke
• 金额: $ 6.85万
• 依托单位: Institute of Life Science, Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557025/
• 关键词: anthrax toxin; GFP; recombinant proteins; 色素性乾皮症; 蛋白質導入

Anthrax toxin is comprised of functionally different two components. Lethal factor (LF) is a effector molecule, while protective antigen (PA) is a molecule which facilitates the ently of LF into cytosol. The N-terminal part of LF molecule is binding domain to PA.Fusion proteins containing the N-terminal part of LF has been shown to be translocated to the cytosol by means of the activity of PA.In this study we attempted to make a recombinant proteins which were translocated into nuclei from outside of cells by interaction with PA.We have constructed the fusion proteins comprising GFP and N-terminal hexsa lysine complex. However, this protein was quite unstable and it did not permit to analyze the translocation of the fusion proteins. Constructions of other form of fusion proteins are in progress.

炭疽毒素由功能不同的两种成分组成。致死因子（LF）是一种效应分子，而保护性抗原（PA）是一种促进LF进入细胞质的分子。 LF 分子的 N 端部分是 PA 的结合域。含有 LF N 端部分的融合蛋白已被证明可以通过 PA 的活性易位到胞质溶胶中。在本研究中，我们尝试制作重组蛋白通过与 PA 相互作用从细胞外易位到细胞核中的蛋白质。我们构建了包含 GFP 和 N 端六角赖氨酸复合物的融合蛋白。然而，这种蛋白质非常不稳定，无法分析融合蛋白的易位。其他形式的融合蛋白的构建正在进行中。

项目成果

Izumi, Y.: "A metalloprotease-disintegrin, MDC9/Meltrin-g/DM9, and PKCd are involved in TPA-induced ecftodomain sheddding of membrane-anchored heparin-binding EGF-like growth factor." EMBO J.17. 7260-7272 (19978)

Izumi, Y.：“金属蛋白酶解整合素、MDC9/Meltrin-g/DM9 和 PKCd 参与 TPA 诱导的膜锚定肝素结合 EGF 样生长因子的外域脱落。”

Umata, T.: "Diphtheria toxin translocation across endosome membrane. A novel cell permeabilization assay reveals new diphtheria toxin fragments in endocytic vesicles." J.Biol.Chem.273. 8351-8359 (1998)

Umata, T.：“白喉毒素跨内体膜易位。一种新型细胞透化测定揭示了内吞囊泡中新的白喉毒素片段。”

Izumi, Y.: "Ametalloprotease-disintegrin, MDC9/Meltrin-g/ADM9, and PKCd are involved in TPA-induced ectodomain shedding of membrane-anchored heparin-binding EGF-like growth factor." EMBO J.17. 7260-7272 (1998)

Izumi, Y.：“Ametalprotease-disintegrin、MDC9/Meltrin-g/ADM9 和 PKCd 参与 TPA 诱导的膜锚定肝素结合 EGF 样生长因子的胞外域脱落。”

岩本 亮: "医学&サイエンスシリーズ 細胞接着のしくみと疾患" 羊土社, 7 (1998)

岩本亮：《医学与科学系列细胞粘附机制与疾病》Yodosha，7 (1998)

Kagawa, T.: "Immune system-related CD9 is expressed in mouse cdentral nervous system myelin at a very late stage of myelination." J.Neurosci. Res.50. 312-320 (1997)

Kakawa, T.：“免疫系统相关的 CD9 在髓鞘形成的最后阶段在小鼠中枢神经系统髓磷脂中表达。”