Sato M, Sato-Miyamoto Y, Kameyama K, Ishikawa N, Imai M, Ito Y, Takagi T
Institute for Protein Research, Osaka University.
J Biochem. 1995 Dec;118(6):1297-302. doi: 10.1093/oxfordjournals.jbchem.a125022.
The internal structure of the recombinant yeast-derived human hepatitis B virus surface antigen vaccine particle was investigated by small-angle neutron scattering using the contrast variation method. Data were collected in aqueous buffer solutions containing 0, 40, 60, and 100% D2O in the q range of 0.005 to 0.2 A-1 at 5 degrees C. The radius of gyration at infinite contrast and the maximum dimension of the particle were estimated to be 107 and 290 A, respectively. The contrast matching point of the particle was determined to correspond to about 30% D2O, indicating that a considerable portion of the vaccine particle is made up of lipids and carbohydrates from yeast. The distance distribution function of the particle at 40% D2O, at which the protein components are matched out to show only the distribution of the remaining lipids and carbohydrates, differed markedly from the functions at other D2Os, and showed the nearly symmetrical profile characteristic of a spherical particle with a diameter of 240 A. The Stuhrmann plot and profile of the distance distribution function at 40% D2O showed that (i) the vaccine is a spherical particle with a diameter of 290 A, in which two different regions in terms of scattering density are distributed radially, (ii) the lipids and carbohydrates form a spherical cluster with a diameter of 240 A in the core region of the particle, and (iii) the surface antigen protein is present in the peripheral region. This architecture of the vaccine particle is favorable for the induction of anti-virus antibodies.
采用对比变化法，通过小角中子散射研究了重组酵母衍生的人乙肝病毒表面抗原疫苗颗粒的内部结构。在5℃下，于0.005至0.2 Å⁻¹的q范围内，在含有0%、40%、60%和100% D₂O的水性缓冲溶液中收集数据。无限对比度下的回转半径和颗粒的最大尺寸估计分别为107 Å和290 Å。确定颗粒的对比匹配点对应于约30% D₂O，这表明疫苗颗粒的相当一部分由酵母中的脂质和碳水化合物组成。在40% D₂O时颗粒的距离分布函数（此时蛋白质成分被匹配掉，仅显示剩余脂质和碳水化合物的分布）与其他D₂O浓度下的函数有显著差异，并且显示出直径为240 Å的球形颗粒的近乎对称的轮廓特征。40% D₂O时的施图尔曼图和距离分布函数轮廓表明：（i）疫苗是直径为290 Å的球形颗粒，其中在散射密度方面有两个不同区域呈径向分布；（ii）脂质和碳水化合物在颗粒的核心区域形成直径为240 Å的球形簇；（iii）表面抗原蛋白存在于周边区域。这种疫苗颗粒结构有利于诱导抗病毒抗体。
