Functionalized Assembly of Proteins by Rational Design of Matrix Bilayr Membrane and Modification of Membrane Targeting Signals of Proteins.

通过合理设计基质双层膜和修饰蛋白质的膜靶向信号来功能化蛋白质组装。

• 批准号: 04455020
• 负责人: KUNITAKE Toyoki
• 金额: $ 1.28万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04455020/
• 关键词: Lipid Bilayr Membrane; Myoglobin; Lipid-Anchoring; Signal Sequence; Membrane-Bound Protein; Protein Assembly; 膜結合性蛋白質; タンパク集積体; 二分子膜; 配向固定化

Directionally controlled anchoring of myoglobin, a water-soluble globular protein, on a dipalmitoylphosphatidylcholine(DPPC) bilayr has been successfully conducted. Lipid-cnchored myoglobin was obtained from a monoalkylated heme derivative and apomyoglobin by cofactor reconstituted method. It was indicated by gel filtration and ultrafiltration studies that the lipid-anchored myoglobin with a hydrophobic alkyl chain, but not native myoglobin and the hydrophilic-anchored myoglobin, is bound to the DPPC bilayr membrane in aqueous dispersion. A cast film of the phospholipid bilayr and the anchored myoglobin displayd anisotropic ESR signals, which depend on the disposition of the cast film in the magnetic field. These results suggest that myoglobin molecules are placed on the lipid bilayr in a fixed orientation by inserting the snchor alkyl chain into the bilayr. The ESR anisotropy was not observed without the anchor.The present research clearly showed that lipid-anchor method is useful for oriented assembly of protein molecules in lipid bilayr membranes.

肌红蛋白是一种水溶性的球状蛋白，在二棕榈酰磷脂酰胆碱（DPPC）双层膜上的定向控制锚定已被成功地进行。以一烷基化血红素衍生物和脱辅基肌红蛋白为原料，采用辅因子重构法制备了脂质化肌红蛋白。凝胶过滤和超滤研究表明，具有疏水烷基链的脂质锚定肌红蛋白，而不是天然肌红蛋白和疏水锚定肌红蛋白，在水性分散体中结合到DPPC双层膜上。磷脂双层和锚定肌红蛋白的流延膜显示各向异性ESR信号，这取决于流延膜在磁场中的布置。这些结果表明，肌红蛋白分子被放置在一个固定的方向上的脂质双层插入的snchor烷基链的双层。在没有锚的情况下，没有观察到ESR各向异性。本研究清楚地表明，脂质-锚方法适用于蛋白质分子在脂质双层膜中的定向组装。

项目成果

三苫 純也: "The Carboxy-terminal 10 amino acid residues of Cytochrome b_5 are necessary for its targeting to the endoplasmic reticulum" The EMBO Journal. 11. 4197-4203 (1992)

Junya Mitoma：“细胞色素 b_5 的羧基末端 10 个氨基酸残基对于其靶向内质网是必需的”The EMBO Journal 11. 4197-4203 (1992)。

浜地 格: "Anisotropic Incorporation of Lipid-Anchored Myoglobin into Phospholipid Bilayer Membrane" Journal of the American Chemical Society. 115. (1993)

Itaru Hamachi：“脂质锚定肌红蛋白各向异性掺入磷脂双层膜”美国化学会杂志 115。（1993）

三苫 純也: "Mitochondrial Targeting of Rat Liver Monoamine Oxidase B is Located at Its Carboxy Terrminus." Journal of Biochemistry. 111. 20-24 (1992)

Junya Mitoma：“大鼠肝脏单胺氧化酶 B 的线粒体靶向位于其羧基末端。”《生物化学杂志》111. 20-24 (1992)

I.Hamachi et al: ""Anisotropic Incorporation of Lipid-Anchored Myoglobin into a Phospholipid Bilayr Membrane."" J.Am. Chem. Soc.115. 1966-4970 (1993)

I.Hamachi 等人：“脂质锚定肌红蛋白各向异性掺入磷脂双层膜。”J.Am.

浜地 格: "Simultaneous Fixation of Orientation of Myoglobin and Metalloporphyrins by Synthetic Bilayer Membrane" Proc.Japan Academy Ser.B. 68. 47-50 (1992)

Itaru Hamachi：“通过合成双层膜同时固定肌红蛋白和金属卟啉的方向”Proc.Japan Academy Ser.B. 68. 47-50 (1992)