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

Abstract

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.