定量荧光共振转移法研究LMP1与脂筏

Biophysical Society of Guang Dong Province Academic Forum - Precise Photons and Life Health Pub Date : 2023-04-04 DOI:10.1117/12.2673642

Zhiwei Wu, Qing Ye, Xiaoyan Wang, Xian-zeng Zhang, S. Xie

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引用次数: 0

摘要

本研究采用三通道显微镜，采用定量荧光共振转移法(E-FRET)研究脂筏破坏对LMP1寡聚化的影响。我们首先用ECFP-only和EYFP-only质粒在我们的成像系统上验证了4个串扰和透流参数以及系统参数G。此外，构建了两个与LMP1相关的基于fret的探针，以研究鼻咽癌细胞系(CNE1)中癌蛋白LMP1的寡聚化。甲基-β-环糊精(m -β cd)用于快速破坏脂筏。FRET图像显示，大部分LMP1低聚物定位于CNE1的内核周膜，脂筏破坏导致LMP1低聚物增强。这些发现为LMP1和脂筏的研究提供了新的视角。

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Study on LMP1 and lipid rafts by quantitative fluorescence resonance transfer method

In this study, three-channel microscopy was used to study the effect of lipid rafts disruption on LMP1 oligomerization by quantitative fluorescence resonance transfer method (E-FRET). We first verified four crosstalk and bleedthrough parameters and the system parameter G on our imaging system by ECFP-only and EYFP-only plasmids. Furtherly, two FRET-based probes associated with LMP1, were constructed to study oligomerization of oncoprotein LMP1 in nasopharyngeal carcinoma cell line (CNE1). Methyl-β-cyclodextrin (MβCD) was used to disrupt lipid rafts, quickly. The FRET images displayed that majority of LMP1 oligomer localized in internal perinuclear membranes of CNE1 and enhancement of LMP1 oligomerization caused by lipid rafts disruption. These findings provided some novel views for LMP1 and lipid rafts.

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Biophysical Society of Guang Dong Province Academic Forum - Precise Photons and Life Health

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