Characterization of the lipid and protein organization in HBsAg viral particles by steady-state and time-resolved fluorescence spectroscopy

被引:33
作者
Greiner, Vanille J. [1 ]
Egele, Caroline [1 ]
Oncul, Sule [1 ]
Ronzon, Frederic [2 ]
Manin, Catherine [2 ]
Klymchenko, Andrey [1 ]
Mely, Yves [1 ]
机构
[1] Univ Strasbourg, UMR CNRS 7213, Lab Biophoton & Pharmacol, Fac Pharm, F-67401 Illkirch Graffenstaden, France
[2] Sanofi Pasteur, F-69280 Marcy Letoile, France
关键词
HBsAg viral particles; Low density lipoprotein; Lipid membranes; Fluorescence spectroscopy; Fluorescent probes; B SURFACE-ANTIGEN; LOW-DENSITY-LIPOPROTEIN; HIV-1 NUCLEOCAPSID PROTEIN; VIRUS-LIKE PARTICLES; HEPATITIS-B; AUSTRALIA-ANTIGEN; NILE-RED; MAXIMUM-ENTROPY; ENERGY-TRANSFER; PROBES;
D O I
10.1016/j.biochi.2010.04.014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hepatitis B surface antigen (HBsAg) particles, produced in the yeast Hansenula polymorpha, are 20 nm particles, composed of S surface viral proteins and host-derived lipids. Since the detailed structure of these particles is still missing, we further characterized them by fluorescence techniques. Fluorescence correlation spectroscopy indicated that the particles are mainly monomeric, with about 70 S proteins per particle. The S proteins were characterized through the intrinsic fluorescence of their thirteen Trp residues. Fluorescence quenching and time-resolved fluorescence experiments suggest the presence of both low emissive embedded Trp residues and more emissive Trp residues at the surface of the HBsAg particles. The low emission of the embedded Trp residues is consistent with their close proximity in alpha-helices. Furthermore. S proteins exhibit restricted movement, as expected from their tight association with lipids. The lipid organization of the particles was studied using viscosity-sensitive DPH-based probes and environment sensitive 3-hydroxyflavone probes, and compared to lipid vesicles and low density lipoproteins (LDLs), taken as models. Like LDLs, the HBsAg particles were found to be composed of an ordered rigid lipid interface, probably organized as a phospholipid monolayer, and a more hydrophobic and fluid inner core, likely composed of triglycerides and free fatty acids. However, the lipid core of HBsAg particles was substantially more polar than the LDL one, probably due to its larger content in proteins and its lower content in sterols. Based on our data, we propose a structural model for HBsAg particles where the S proteins deeply penetrate into the lipid core. (C) 2010 Elsevier Masson SAS. All rights reserved.
引用
收藏
页码:994 / 1002
页数:9
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