Barklis E, McDermott J, Wilkens S, Fuller S, Thompson D
Vollum Institute and Department of Microbiology, Oregon Health Sciences University, Portland, Oregon 97201-3098, USA.
J Biol Chem. 1998 Mar 27;273(13):7177-80. doi: 10.1074/jbc.273.13.7177.
In an in vitro system that mimics the assembly of immature human immunodeficiency virus (HIV) particles, ordered arrays of HIV-1 capsid (CA) proteins encoded by the viral gag gene have been obtained by incubation of histidine-tagged capsid proteins (His-HIVCA) beneath lipid monolayers containing the nickel-chelating lipid, 1,2-di-O-hexadecyl-sn-glycero-3-(1'-2"-R-hydroxy-3'-N-(5-amino-1- carboxypentyl)iminodiacetic acid)propyl ether. The membrane-bound His-HIVCA proteins formed small crystalline arrays of primitive (p1) unit cells with dimensions of a = 74.2 A, b = 126.2 A, gamma = 89.3 degrees. The image-analyzed two-dimensional projection of His-HIVCA assemblies shows a cage-like lattice, consisting of hexamer and trimer units, surrounding protein-free cage holes. The hexamer-coordinated cage holes of 26.3-A diameter are spaced at 74. 2-A intervals: these distances, and the hexamer-trimer arrangement, are consistent with previous, lower resolution studies on immature HIV-1 virus particles produced in vivo. Additionally, HIV-1 matrix protein trimer unit structures align to the His-HIVCA trimer units such that residues previously shown to interact with the HIV-1 gp120/gp41 envelope protein complex are oriented toward the hexamer cage holes. Our results form a bridge between results from conventional methods for the analysis of HIV particle structure.
在一个模拟未成熟人类免疫缺陷病毒（HIV）颗粒组装的体外系统中，通过在含有镍螯合脂质1,2 - 二 - O - 十六烷基 - sn - 甘油 - 3 -（1'-2"-R - 羟基 - 3'-N -（5 - 氨基 - 1 - 羧基戊基）亚氨基二乙酸）丙醚的脂质单层下方孵育组氨酸标记的衣壳蛋白（His - HIVCA），获得了由病毒gag基因编码的HIV - 1衣壳（CA）蛋白的有序阵列。膜结合的His - HIVCA蛋白形成了原始（p1）晶胞的小晶体阵列，其尺寸为a = 74.2 Å，b = 126.2 Å，γ = 89.3°。His - HIVCA组装体的图像分析二维投影显示出一个笼状晶格，由六聚体和三聚体单元组成，围绕着无蛋白的笼状孔。直径为26.3 Å的六聚体配位笼状孔以74.2 Å的间隔排列：这些距离以及六聚体 - 三聚体排列与先前对体内产生的未成熟HIV - 1病毒颗粒的低分辨率研究一致。此外，HIV - 1基质蛋白三聚体单元结构与His - HIVCA三聚体单元对齐，使得先前显示与HIV - 1 gp120/gp41包膜蛋白复合物相互作用的残基朝向六聚体笼状孔。我们的结果在用于分析HIV颗粒结构的传统方法的结果之间架起了一座桥梁。
