MacDonald R I
Biochemistry. 1985 Jul 16;24(15):4058-66. doi: 10.1021/bi00336a039.
To determine whether polyethylene glycol (PEG) causes growth of liposomes by affecting them directly or indirectly, vesicles composed of phosphatidylcholine were exposed to increasing concentrations of Mr 15 000-20 000 PEG or Mr 40 000 dextran either by direct mixing or across a dialysis membrane. After incubation at room temperature and dilution below at least 5% (w/w) polymer, the vesicles were monitored for fluorescence energy transfer and for absorbance at 400 nm. PEG induced the same levels of dequenching or lipid mixing and increased turbidity, regardless of whether the vesicles had been mixed directly with or dialyzed against PEG. These changes occurred within 5-15 min of polymer application. It is concluded that the increased lipid mixing and/or increased turbidity, indicating vesicle growth, resulted from an indirect effect of PEG on the vesicles--most likely dehydration. Dextran, in contrast to PEG, induced less dequenching and/or less turbidity increase when vesicles were directly mixed with, as opposed to dialyzed against, dextran. Although dextran not in contact with vesicles and with osmotic activity comparable to PEG was able to cause a degree of membrane fusion similar to that of PEG, therefore, the dehydrating effect of dextran could be mitigated if it were allowed to interact with vesicles. In further support of membrane dehydration as a precursor to membrane fusion, lipid mixing among sonicated and sonicated, frozen-thawed vesicles dialyzed against sucrose increased as a function of sucrose concentration. Vesicle morphology generally determined the maximal degree of membrane fusion inducible by the polymers.(ABSTRACT TRUNCATED AT 250 WORDS)
为了确定聚乙二醇(PEG)是直接还是间接影响脂质体从而使其生长,通过直接混合或透过透析膜,将由磷脂酰胆碱组成的囊泡暴露于浓度不断增加的分子量为15000 - 20000的PEG或分子量为40000的葡聚糖中。在室温下孵育并稀释至聚合物浓度至少低于5%(w/w)后,监测囊泡的荧光能量转移和400nm处的吸光度。无论囊泡是直接与PEG混合还是与PEG进行透析,PEG都能诱导相同程度的去淬灭或脂质混合,并增加浊度。这些变化在施加聚合物后的5 - 15分钟内发生。得出的结论是,脂质混合增加和/或浊度增加表明囊泡生长,这是PEG对囊泡的间接作用导致的——最有可能是脱水作用。相比之下,当囊泡与葡聚糖直接混合而非进行透析时,葡聚糖诱导的去淬灭和/或浊度增加较少。尽管与PEG渗透压相当但不与囊泡接触的葡聚糖能够引起与PEG程度相似的膜融合,然而,如果允许葡聚糖与囊泡相互作用,其脱水作用可能会减弱。为进一步支持膜脱水是膜融合的前奏这一观点,对经超声处理以及超声处理后再冻融的囊泡进行针对蔗糖的透析,脂质混合随蔗糖浓度增加。囊泡形态通常决定了聚合物可诱导的最大膜融合程度。(摘要截短于250字)