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首页> 外文期刊>Biochemistry >Pyrene Cholesterol Reports the Transient Appearance of Nonlamellar Intermediate Structures during Fusion of Model Membranes
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Pyrene Cholesterol Reports the Transient Appearance of Nonlamellar Intermediate Structures during Fusion of Model Membranes

机译：yr胆固醇报告模型膜融合过程中非薄片中间结构的瞬态出现。

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We have hypothesized that modulating the free energy of hydrophobic mismatch (HM) might be a principal means to control the fusion process and that is may be a role of cholesterol to counteract HM and make membranes fusogenic. To test these hypotheses, we examined the ability of cholesterol 1-phrenebutyrate (PY-Ch) and other pyrene-containing fluorescent probes to report interstices formed during the L_(#alpha#)-H_(II) transition of DiPoPE in terms of changes in excimer/monomer (E?M) fluorescence ratios. We found a significant (>150%) increase in the PY-Ch E/M in the hexagonal phase relative to the lamellar phase, presumably resulting from redistribution of PY-Ch from the curved lamellar leaflets to coexisting HMs that constitute 20 vol% of this phase. Al other probes showed a much smaller or even an opposite (PY-hexadecanoic acid) effect. The time course of the PY-Ch E/M ratio during fusion of DOPC/PE/Ch small unilamellar vesicles showed a transient increase with a subsequent decrease, consistent with fusion proceeding through intermediates with significant HM. The amplitude and position of the maximum in E/M correalted with the rate of contents mixing. An increase in E/M was not seen when lipid mixing occurred in the absence of contents mixing. Our results suggest that pY-Ch provides a tool for monitoring fusion intermediates that occur after the initial fusion intermediate but prior to pore formation, possibly by accumulating in regins associated with HM.
机译：我们假设调节疏水性错配（HM）的自由能可能是控制融合过程的主要手段，并且可能是胆固醇抵消HM并使膜融合的作用。为了检验这些假设，我们检查了胆固醇1-苯丁酸酯（PY-Ch）和其他含pyr的荧光探针报告DiPoPE在L _（＃alpha＃）-H_（II）过渡期间形成的空隙的变化能力。在准分子/单体（E2M）荧光比方面。我们发现六角形相中的PY-Ch E / M相对于层状相显着增加（> 150％），这可能是由于PY-Ch从弯曲层状小叶重新分布到共存的HM所致，而HMs占层状体积的20％。这个阶段。所有其他探针显示出更小的甚至相反的（PY-十六烷酸）作用。在DOPC / PE / Ch小单层囊泡融合过程中，PY-Ch E / M比的时间过程显示出瞬时增加，随后减少，这与通过具有显着HM的中间体进行融合相一致。 E / M最大值的振幅和位置与内容物混合速率相关。当在没有内容物混合的情况下进行脂质混合时，未观察到E / M的增加。我们的结果表明，pY-Ch提供了一种监测融合中间体的工具，该融合中间体可能发生在初始融合中间体之后但在孔形成之前，可能是通过与HM相关的蛋白积累而引起的。

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