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通过Langmuir单分子层研究对甘草酸与脂质筏模型的界面行为进行研究。

Investigation of interfacial behavior of glycyrrhizin with a lipid raft model via a Langmuir monolayer study.

作者信息

Sakamoto Seiichi, Nakahara Hiromichi, Uto Takuhiro, Shoyama Yukihiro, Shibata Osamu

机构信息

Department of Biophysical Chemistry, Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch, Sasebo, Nagasaki 859-3298, Japan.

出版信息

Biochim Biophys Acta. 2013 Apr;1828(4):1271-83. doi: 10.1016/j.bbamem.2013.01.006. Epub 2013 Jan 16.

Abstract

An interaction of glycyrrhizin (GC) with a lipid raft biomembrane model that consisted of N-palmitoyl-d-erythro-sphingosylphosphorylcholine (PSM), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and cholesterol (CHOL) was systematically studied using the Langmuir monolayer technique. To construct the lipid raft model, the surface pressure (π)-molecular area (A) and surface potential (ΔV)-A isotherms for three-component (PSM/DOPC/CHOL) systems on 0.02M Tris buffer with 0.13M NaCl (pH7.4) were primarily measured by changing their compositions. Thermodynamic and interaction parameters for binary PSM/DOPC and PSM/CHOL systems revealed that PSM interacts more strongly with CHOL than with DOPC. In addition, a morphological analysis performed with Brewster angle microscopy (BAM) and fluorescence microscopy (FM) revealed an optimal ratio of PSM/DOPC/CHOL (1/1/1, by mole) as a model of lipid rafts. Second, the interaction of GC with the ternary PSM/DOPC/CHOL monolayers was investigated on Tris buffer solutions containing different GC concentrations (1, 5, 10, 25, and 50μM). In BAM and FM images, microdomains were found to become smaller by increasing the GC concentration in the subphase, suggesting that GC regulates the size of raft domains, which provide dynamic scaffolding for numerous cellular processes. More interestingly, the distinctive GC striped regions were formed at the interface at 50μM, which shows that GC divides the ternary monolayer into pieces. This phenomenon was observed only in the presence of CHOL in the monolayer. These results suggest that CHOL plays an essential role in the interaction with GC, which results in one of the major activities associated with saponins' membrane disruption.

摘要

利用朗缪尔单分子层技术,系统研究了甘草酸(GC)与由N-棕榈酰-d-赤藓糖基神经鞘磷脂酰胆碱(PSM)、1,2-二油酰基-sn-甘油-3-磷酸胆碱(DOPC)和胆固醇(CHOL)组成的脂筏生物膜模型之间的相互作用。为构建脂筏模型,首先通过改变其组成,测量了在含0.13M NaCl(pH7.4)的0.02M Tris缓冲液中三组分(PSM/DOPC/CHOL)体系的表面压力(π)-分子面积(A)等温线和表面电位(ΔV)-A等温线。二元PSM/DOPC和PSM/CHOL体系的热力学和相互作用参数表明,PSM与CHOL的相互作用比与DOPC的更强。此外,用布鲁斯特角显微镜(BAM)和荧光显微镜(FM)进行的形态分析表明,PSM/DOPC/CHOL(摩尔比1/1/1)是脂筏的最佳模型比例。其次,在含有不同GC浓度(1、5、10、25和50μM)的Tris缓冲溶液中,研究了GC与三元PSM/DOPC/CHOL单分子层的相互作用。在BAM和FM图像中,发现通过增加亚相中GC的浓度,微区会变小,这表明GC调节了筏结构域的大小,而筏结构域为众多细胞过程提供动态支架。更有趣的是,在50μM时在界面处形成了独特的GC条纹区域,这表明GC将三元单分子层分成了小块。这种现象仅在单分子层中存在CHOL时才观察到。这些结果表明,CHOL在与GC的相互作用中起重要作用,这导致了与皂苷膜破坏相关的主要活性之一。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff85/7089610/57e28ddc99e1/fx1_lrg.jpg

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