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

在二棕榈酰磷脂酰胆碱(DPPC)双分子膜上成功地实现了水溶性球状蛋白肌红蛋白的定向锚定。以一种单烷基化的血红素衍生物和去肌红蛋白为原料，通过辅因子重组法制备了脂质修饰肌红蛋白。凝胶过滤和超滤研究表明，具有疏水性烷基链的脂锚型肌红蛋白，而不是天然肌红蛋白和亲水性锚定肌红蛋白，在水分散体系中结合在DPPC双层膜上。磷脂双层和锚定的肌红蛋白的铸膜层显示各向异性的ESR信号，这取决于铸膜层在磁场中的配置。这些结果表明，肌红蛋白分子通过将短链烷基链插入到脂质双分子层中而以固定的方向放置在脂质双分子层上。目前的研究表明，脂类锚定方法对于蛋白质分子在脂膜中的定向组装是有用的。

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.