Baeza I, Wong C, Mondragón R, González S, Ibáñez M, Farfán N, Argüello C
Departmento de Bioquímica, Escuela Nacional de Ciencias Biológicas del IPN, Mexico, D.F.
J Mol Evol. 1994 Dec;39(6):560-8. doi: 10.1007/BF00160401.
Liposomes formed from egg-yolk phosphatidylcholine:egg-yolk phosphatidate (molar ratio 2:1) containing pBR322 DNA and DNase I were induced to form, with divalent cations, bilayer/nonbilayer phase transitions of phosphatidate which allowed cation diffusion into liposomes; then cation diffusion was measured by the activation of the hydrolysis of DNase I on DNA. The formation of phosphatidate transitions on liposomes was demonstrated by freeze-fracture and 31P NMR, and a direct correlation between the formation of phosphatidate transitions and the transbilayer diffusion of cations was found: only Ca2+ and Mn2+, which induce phase transitions, were able to penetrate liposomes and triggered the DNase I activity; in addition, Ca2+ at higher concentrations (10 mM) caused fusion of liposomes, whereas Mn2+ did not, suggesting that transitions induced by Mn2+ participated only in the diffusion of this ion; furthermore, Mg2+ neither formed phase transitions nor triggered the enzymatic activity. The liposomes studied represent more dynamic structures that can form phosphatidate structures involved in both (1) the interchange of divalent cations with the surroundings, thereby modulating encapsulated enzymes, and (2) the fusion of lipid vesicles probably implicated in the enrichment of liposomal content in the early Precambian Earth.
由蛋黄磷脂酰胆碱与蛋黄磷脂酸(摩尔比为2:1)形成的脂质体,含有pBR322 DNA和DNA酶I,在二价阳离子作用下诱导形成磷脂酸的双层/非双层相变,这使得阳离子能够扩散到脂质体中;然后通过DNA酶I对DNA水解的激活来测量阳离子扩散。通过冷冻蚀刻和31P核磁共振证实了脂质体上磷脂酸相变的形成,并且发现磷脂酸相变的形成与阳离子的跨膜扩散之间存在直接相关性:只有能诱导相变的Ca2+和Mn2+能够穿透脂质体并触发DNA酶I的活性;此外,较高浓度(10 mM)的Ca2+会导致脂质体融合,而Mn2+则不会,这表明由Mn2+诱导的相变仅参与该离子的扩散;此外,Mg2+既不形成相变也不触发酶活性。所研究的脂质体代表了更具动态性的结构,这些结构能够形成参与以下两方面的磷脂酸结构:(1)二价阳离子与周围环境的交换,从而调节包封的酶;(2)脂质囊泡的融合,这可能与前寒武纪早期地球脂质体内容物的富集有关。