Laboratorio di Biofisica Molecolare and UdR CNISM, Dipartimento di Fisica, Università della Calabria, Rende (CS), Italy.
Eur Biophys J. 2010 May;39(6):921-7. doi: 10.1007/s00249-009-0442-0. Epub 2009 Apr 7.
Electron spin resonance (ESR) spectroscopy is used to study the transfer of stearic acids between human serum albumin (HSA) and sterically stabilized liposomes (SSL) composed of dipalmitoylphosphatidylcholine (DPPC) and of submicellar content of poly(ethylene glycol:2000)-dipalmitoylphosphatidylethanolamine (PEG:2000-DPPE). Protein/lipid dispersions are considered in which spin-labelled stearic acids at the 16th carbon atom along the acyl chain (16-SASL) are inserted either in the protein or in the SSL. Two component ESR spectra with different rotational mobility are obtained over a broad range of temperature and membrane composition. Indeed, superimposed to an anisotropic protein-signal, appears a more isotropic lipid-signal. Since in the samples only one matrix (protein or membranes) is spin-labelled, the other component accounts for the transfer of 16-SASL between albumin and membranes. The two components have been resolved and quantified by spectral subtractions, and the fraction, f (p) (16-SASL), of spin labels bound non-covalently to the protein has been used to monitor the transfer. It is found that it depends on the type of donor and acceptor matrix, on the physical state of the membranes and on the grafting density of the polymer-lipids. Indeed, it is favoured from SSL to HSA and the fraction of stearic acids transferred increases with temperature in both directions of transfer. Moreover, in the presence of polymer-lipids, the transfer from HSA to SSL is slightly attenuated, especially in the brush regime of the polymer-chains. Instead, the transfer from SSL to HSA is favoured by the polymer-lipids much more in the mushroom than in the brush regime.
电子自旋共振(ESR)光谱用于研究硬脂酸在人血清白蛋白(HSA)和由二棕榈酰磷脂酰胆碱(DPPC)和亚胶束含量的聚乙二醇:2000-二棕榈酰磷脂酰乙醇胺(PEG:2000-DPPE)组成的刚性稳定脂质体(SSL)之间的转移。研究了蛋白质/脂质分散体,其中在酰链第 16 个碳原子处插入了带有自旋标记的硬脂酸(16-SASL),要么插入蛋白质中,要么插入 SSL 中。在较宽的温度和膜组成范围内,获得了具有不同旋转流动性的两个组分 ESR 谱。实际上,在各向异性的蛋白质信号上叠加了更各向同性的脂质信号。由于在样品中只有一个基质(蛋白质或膜)被自旋标记,因此其他组分则代表 16-SASL 在白蛋白和膜之间的转移。通过光谱减法对两个组分进行了分辨和定量,并且使用非共价结合到蛋白质上的自旋标记的分数 f(p)(16-SASL)来监测转移。发现它取决于供体和受体基质的类型,膜的物理状态以及聚合物脂质的接枝密度。实际上,它有利于从 SSL 到 HSA,并且在两个转移方向上,硬脂酸的转移分数随温度增加而增加。此外,在聚合物脂质存在的情况下,从 HSA 到 SSL 的转移略有减弱,尤其是在聚合物链的刷状区。相反,聚合物脂质更有利于在蘑菇区而非刷状区从 SSL 到 HSA 的转移。