MEMBRANE PERMEABILIZATION BY DIFFERENT REGIONS OF THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 TRANSMEMBRANE GLYCOPROTEIN GP41
MEMBRANE PERMEABILIZATION BY DIFFERENT REGIONS OF THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 TRANSMEMBRANE GLYCOPROTEIN GP41
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DOI：
10.1128/jvi.69.7.4095-4102.1995
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发表时间：
1995-07-01
影响因子：
5.4
通讯作者：
CARRASCO, L
中科院分区：
文献类型：
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作者：
ARROYO, J;BOCETA, M;CARRASCO, L
人类免疫缺陷病毒1型（HIV-1）的跨膜糖蛋白（gp41）与HIV感染期间观察到的细胞病理学有关。位于氨基末端的第一个氨基酸参与了膜融合和合胞体的形成，而位于羧基末端的序列被预测与膜相互作用并改变膜的渗透性。克隆了HIV-1 gp41基因，并利用pET载体在大肠杆菌细胞中表达，分析了该蛋白产生的膜通透性变化。该系统非常适合于以诱导方式表达毒性基因和分析修饰膜通透性的蛋白质的功能。尽管gp41蛋白的表达水平较低，但它能增强细菌膜对潮霉素B的通透性。为了定位gp41负责这些作用的区域，在大肠杆菌中诱导表达了gp41不同部分的许多片段。gp41的两个区域被证明可以增加膜通透性：一个位于羧基端，在那里预测了两个高度二位螺旋，另一个区域对应于膜跨越区域。包含该结构域的gp41中心区域的表达对大肠杆菌细胞具有高度裂解性，并增加了细胞膜对许多化合物的通透性。根据hiv诱导的细胞病理学和gp41结构对这些发现进行了讨论。
The transmembrane glycoprotein (gp41) of human immunodeficiency virus type 1 (HIV-1) has been implicated in the cytopathology observed during HIV infection. The first amino acids located at the amino terminus are involved in membrane fusion and syncytium formation, while sequences located at the carboxy terminus have been predicted to interact with membranes and modify membrane permeability. The HIV-1 gp41 gene has been cloned and expressed in Escherichia coli cells by using pET vectors to analyze changes in membrane permeability produced by this protein. This system is well suited for expressing toxic genes in an inducible manner and for analyzing the function of proteins that modify membrane permeability. gp41 enhances the permeability of the bacterial membrane to hygromycin B despite the low level of expression of this protein. To localize the regions of gp41 responsible for these effects, a number of fragments spanning different portions of gp41 were inducibly expressed in E. coli. Two regions of gp41 were shown to increase membrane permeability: one located at the carboxy terminus, where two highly amphipatic helices have been predicted, and another one corresponding to the membrane-spanning domain. Expression of the central region of gp41 comprising this domain was highly lytic for E. coli cells and increased membrane permeability to a number of compounds. These findings are discussed in the light of HIV-induced cytopathology and gp41 structure.