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

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。

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.