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荧光寿命成像监测的肽-膜相互作用:转导肽 10 的研究案例。

Peptide-Membrane Interactions Monitored by Fluorescence Lifetime Imaging: A Study Case of Transportan 10.

机构信息

Dipartimento di Fisica e Chimica-Emilio Segré, Università degli Studi di Palermo, Viale delle Scienze ed. 18 90128, Palermo, Italy.

Department of Pharmacy, University of Copenhagen, Universitetsparken 2 2100, Copenhagen, Denmark.

出版信息

Langmuir. 2021 Nov 9;37(44):13148-13159. doi: 10.1021/acs.langmuir.1c02392. Epub 2021 Oct 29.

Abstract

The interest on detailed analysis of peptide-membrane interactions is of great interest in both fundamental and applied sciences as these may relate to both functional and pathogenic events. Such interactions are highly dynamic and spatially heterogeneous, making the investigation of the associated phenomena highly complex. The specific properties of membranes and peptide structural details, together with environmental conditions, may determine different events at the membrane interface, which will drive the fate of the peptide-membrane system. Here, we use an experimental approach based on the combination of spectroscopy and fluorescence microscopy methods to characterize the interactions of the multifunctional amphiphilic peptide transportan 10 with model membranes. Our approach, based on the use of suitable fluorescence reporters, exploits the advantages of phasor plot analysis of fluorescence lifetime imaging microscopy measurements to highlight the molecular details of occurring membrane alterations in terms of rigidity and hydration. Simultaneously, it allows following dynamic events in real time without sample manipulation distinguishing, with high spatial resolution, whether the peptide is adsorbed to or inserted in the membrane.

摘要

对肽-膜相互作用进行详细分析的兴趣在基础科学和应用科学中都非常大,因为这些相互作用可能与功能和病理事件有关。这些相互作用具有高度动态和空间异质性,使得相关现象的研究非常复杂。膜的特定性质和肽结构细节,以及环境条件,可能决定膜界面上的不同事件,从而决定肽-膜系统的命运。在这里,我们使用一种基于光谱学和荧光显微镜方法相结合的实验方法来表征多功能两亲肽转运蛋白 10 与模型膜的相互作用。我们的方法基于使用合适的荧光报告物,利用荧光寿命成像显微镜测量的相图分析的优势来突出发生的膜变化在刚性和水合方面的分子细节。同时,它允许在不进行样品处理的情况下实时跟踪动态事件,以高空间分辨率区分肽是吸附在膜上还是插入膜中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18fc/8582253/87e6da6fc432/la1c02392_0002.jpg

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