Recombinant subviral particles from tick-borne encephalitis virus are fusogenic and provide a model system for studying flavivirus envelope glycoprotein functions
Recombinant subviral particles from tick-borne encephalitis virus are fusogenic and provide a model system for studying flavivirus envelope glycoprotein functions
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DOI：
10.1128/jvi.70.7.4549-4557.1996
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发表时间：
1996-07-01
影响因子：
5.4
通讯作者：
Heinz, FX
中科院分区：
文献类型：
--
作者：
Schalich, J;Allison, SL;Heinz, FX
通过在 COS 细胞中共表达蜱传脑炎病毒的包膜 (E) 和前膜 (prM) 蛋白获得的重组亚病毒颗粒 (RSP) (S. L. Allison、K. Stadler、C. W. Mandl、C. Kunz 和 F. X. Heinz, J. Virol. 69:5816-5820, 1995) 被广泛表征并显示为有序的含有包膜糖蛋白的结构，其结构和功能特性与病毒颗粒包膜中的非常相似。该颗粒呈球形，直径约30 nm，在蔗糖梯度中的浮力密度为1.14 g/cm(3)。它们含有具有内切糖苷酶H抗性聚糖的成熟E蛋白以及完全裂解的成熟M蛋白。 prM 的裂解需要胞吐室中的酸性 pH 值，可以通过用氯化铵处理转染细胞来抑制，这意味着 RSP 和病毒体有共同的成熟途径，RSP 掺入了 [C-14] 胆碱，但不掺入 [H-3] 尿苷，表明它们含有脂质但可能缺乏核酸。与病毒粒子相比，RSP 的包膜蛋白表现出天然抗原和寡聚结构，并且在酸性 pH 值（pH
Recombinant subviral particles (RSPs) obtained by coexpression of the envelope (E) and premembrane (prM) proteins of tick-borne encephalitis virus in COS cells (S. L. Allison, K. Stadler, C. W. Mandl, C. Kunz, and F. X. Heinz, J. Virol. 69:5816-5820, 1995) were extensively characterized and shown to be ordered structures containing envelope glycoproteins with structural and functional properties very similar to those in the virion envelope. The particles were spherical, with a diameter of about 30 nm and a buoyant density of 1.14 g/cm(3) in sucrose gradients, They contained mature E proteins with endoglycosidase H-resistant glycans as well as fully cleaved mature M proteins. Cleavage of prM, which requires an acidic pH in exocytic compartments, could be inhibited by treatment of transfected cells with ammonium chloride, implying a common maturation pathway for RSPs and virions, RSPs incorporated [C-14]choline but not [H-3]uridine, demonstrating that they contain lipid but probably lack nucleic acid. The envelope proteins of RSPs exhibited a native antigenic and oligomeric structure compared with virions, and incubation at an acidic pH (pH