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一个用于研究生物电活性分子与脂质纳米域相互作用的仿生平台。

A biomimetic platform to study the interactions of bioelectroactive molecules with lipid nanodomains.

作者信息

Marquês Joaquim T, Viana Ana S, de Almeida Rodrigo F M

机构信息

Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa , Ed. C8, Campo Grande, 1749-016 Lisboa, Portugal.

出版信息

Langmuir. 2014 Oct 28;30(42):12627-37. doi: 10.1021/la503086a. Epub 2014 Oct 16.

Abstract

In this work, we developed a biomimetic platform where the study of membrane associated redox processes and high-resolution imaging of lipid nanodomains can be both performed, based on a new functional gold modification, l-cysteine self-assembled monolayer. This monolayer proved to be ideal for the preparation of defect-free planar supported lipid bilayers (SLBs) where nanodomains with height difference of ∼1.5 nm are clearly resolved by atomic force microscopy. Single and multicomponent lipid compositions were used, leading to the formation of different phases and domains mimicking the lateral organization of cellular membranes, and in all cases stable and continuous bilayers were obtained. These platforms were tested toward the interaction with bioelectroactive molecules, the antioxidant quercetin, and the hormone epinephrine. Despite the weak interaction detected between epinephrine and lipid bilayers, our biomimetic interface was able to sense the redox process of membrane-bound epinephrine, obtain its surface concentration (9.36 × 10(-11) mol/cm(2) for a fluid bilayer), and estimate a mole fraction membrane/water partition coefficient (Kp) from cyclic voltammetric measurements (1.13 × 10(4) for a fluid phase membrane). This Kp could be used to quantitatively describe the minute changes observed in the photophysical properties of epinephrine intrinsic fluorescence upon its interaction with liposome suspensions. Moreover, we showed that the lipid membrane stabilizes epinephrine structure, preventing its oxidation, which occurs in neutral aqueous solution, and that epinephrine partition and mobility in membranes depends on lipid phase, expanding our knowledge on hormone membrane interactions.

摘要

在这项工作中,我们基于一种新的功能性金修饰——L-半胱氨酸自组装单分子层,开发了一个仿生平台,在该平台上可以同时进行膜相关氧化还原过程的研究以及脂质纳米域的高分辨率成像。事实证明,这种单分子层非常适合制备无缺陷的平面支撑脂质双层(SLB),通过原子力显微镜可以清晰分辨出高度差约为1.5 nm的纳米域。使用了单组分和多组分脂质组合物,导致形成了模仿细胞膜横向组织的不同相和域,并且在所有情况下都获得了稳定且连续的双层。这些平台针对与生物电活性分子、抗氧化剂槲皮素和激素肾上腺素的相互作用进行了测试。尽管检测到肾上腺素与脂质双层之间的相互作用较弱,但我们的仿生界面能够感知膜结合肾上腺素的氧化还原过程,获得其表面浓度(对于流体双层为9.36×10⁻¹¹ mol/cm²),并通过循环伏安测量估算摩尔分数膜/水分配系数(Kp)(对于流体相膜为1.13×10⁴)。这个Kp可用于定量描述肾上腺素固有荧光在与脂质体悬浮液相互作用时光物理性质中观察到的微小变化。此外,我们表明脂质膜稳定了肾上腺素的结构,防止其在中性水溶液中发生氧化,并且肾上腺素在膜中的分配和流动性取决于脂质相,这扩展了我们对激素膜相互作用的认识。

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