Hepatitis B subvirus particles display both a fluid bilayer membrane and a strong resistance to freeze drying: a study by solid‐state NMR, light scattering, and cryo‐electron microscopy/tomography

Article Article scientifique Article Hepatitis B subvirus particles display both a fluid bilayer membrane and a strong resistance to freeze drying: a study by solid‐state NMR, light scattering, and cryo‐electron microscopy/tomography eng 2013-07-09 2013-07-09 author Grélard Axelle Institute of Chemistry and Biology of Membranes and NanoobjectsUniversity of BordeauxChemistry and Biology of Membranes and Nanoobjects (CBMN)Unité Mixte de Recherche (UMR) 5248PessacFrance author Guichard Paul Institut CurieCentre de RechercheOrsayFrance 845268 Département de biologie moléculaire et cellulaire (MOCEL) author Bonnafous Pierre Institute of Chemistry and Biology of Membranes and NanoobjectsUniversity of BordeauxChemistry and Biology of Membranes and Nanoobjects (CBMN)Unité Mixte de Recherche (UMR) 5248PessacFrance author Marco Sergio Institut CurieCentre de RechercheOrsayFrance author Lambert Luc-Olivier Institute of Chemistry and Biology of Membranes and NanoobjectsUniversity of BordeauxChemistry and Biology of Membranes and Nanoobjects (CBMN)Unité Mixte de Recherche (UMR) 5248PessacFrance 87720 author Manin Catherine Sanofi PasteurMarcy l'ÉtoileFrance author Ronzon Frédéric Sanofi PasteurMarcy l'ÉtoileFrance author Dufourc Erick J. Institute of Chemistry and Biology of Membranes and NanoobjectsUniversity of BordeauxChemistry and Biology of Membranes and Nanoobjects (CBMN)Unité Mixte de Recherche (UMR) 5248PessacFrance 4316-4326 10.1096/fj.13-232843 23839934 0892-6638 https://onlinelibrary.wiley.com/doi/abs/10.1096/fj.13-232843 <p>Hepatitis B surface antigen (HBsAg) subvirus particles produced from yeast share immunological determinants with mature viruses, which enable the use of HBsAg as a potent antigen for human vaccination. Because the intimate structure of such pseudoviral particles is still a matter of debate, we investigated the robustness of the external barrier and its structure and dynamics using the noninvasive solid- state NMR technique. This barrier is made of 60% proteins and 40% lipids. Phospholipids represent 83% of all lipids, and chain unsaturation is of 72%. Dynamics was reported by embedding small amounts of deuterium chain-labeled unsaturated phospholipid into the external barrier of entire subviral particles, while controlling particle integrity by cryoelectron microscopy, tomography, and light scattering. Variable preparation modes were used, from mild incubation of small unila-mellar vesicles to very stringent incorporation with freeze-drying. A lipid bilayer structure of 4- to 5-nm thickness was evidenced with a higher rigidity than that of synthetic phospholipid vesicles, but nonetheless reflecting a fluid membrane (50 –52% of maximum rigidity) in agreement with the elevated unsaturation content. The HBsAg particles of 20- to 24-nm diameter were surprisingly found resistant to lyophilization, in such a way that trapped water inside particles could not be removed. These dual properties bring more insight into the mode of action of native subviral particles and their recombinant counterparts used in vaccines.</p> Deuterium labeling Lipid incorporation Lipid thickness Order parameter Trapped water true The FASEB journal 27 10 83cf63e1-961c-42fe-a28b-b03cacb3a9b8 Article (Published version) Restricted LAMBERT01.pdf 363368 application/pdf 43337e91557c873dedc949a9fdec27cadb2cb77c56bbc1adb43258f314d4aae3 b918a8ed-5fac-4a52-933e-027809083328 Supplemental data Public fsb2027010032-sup-0001-2.pdf 1573236 application/pdf 01d68c1897ee20e887f26b9cd16afcfcb290cc4a82cf936957fa57cdc9c8414c restrictedAccess

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Hepatitis B subvirus particles display both a fluid bilayer membrane and a strong resistance to freeze drying: a study by solid‐state NMR, light scattering, and cryo‐electron microscopy/tomography	The FASEB journal			2013	72	0

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