Preferential recognition of monomeric CCR5 expressed in cultured cells by the HIV-1 envelope glycoprotein gp120 for the entry of R5 HIV-1
Preferential recognition of monomeric CCR5 expressed in cultured cells by the HIV-1 envelope glycoprotein gp120 for the entry of R5 HIV-1
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HIV-1包膜糖蛋白gp120优先识别培养细胞中表达的单体CCR5，以促进R5 HIV-1的进入
DOI：
10.1016/j.virol.2013.12.034
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Maeda Y
中科院分区：
文献类型：
--
作者：
Nakano Y;Monde K;Terasawa H;Yuan Y;Yusa K;Harada S;Maeda Y
双分子荧光互补（BiFC）和蛋白质印迹分析表明，CCR 5以组成型同源寡聚体的形式存在，其拮抗剂如马拉韦罗（MVC）和TAK-779进一步增强了CCR 5。通过识别CCR 5的不同表位的单克隆抗体的染色显示CCR 5寡聚体在结构上不同于单体。为了确定CCR 5的哪些形式被CCR 5很好地识别，使用HIV-I进入，通过荧光激活细胞分选仪收集CD 4阳性293 T细胞中的BiFC阳性和阴性细胞级分，并使用包括R5和R5 X4的Env用CCR 5假型化的报道基因HIV-I感染。R5和双R5 HIV-1基本上感染BiFC阴性组分而不是BiFC阳性组分，表明R5和双R5 Env优先识别单体CCR 5。虽然CCR 5拮抗剂增强CCR 5的寡聚化，但发现抗MVC的HIV-1仍然识别MVC结合和未结合形式的单体CCR 5，这表明R5 HIV-1对单体CCR 5的限制使用。
Bimolecular fluorescence complementation (BiFC) and western blot analysis demonstrated that CCR5 exists as constitutive homo-oligomers, which was further enhanced by its antagonists such as maraviroc (MVC) and TAK-779. Staining by monoclonal antibodies recognizing different epitopes of CCR5 revealed that CCR5 oligomer was structurally different from the monomer. To determine which forms of CCR5 are well recognized by CCR5-using HIV-1 for the entry, BiFC-positive and -negative cell fractions in CD4-positive 293T cells were collected by fluorescent-activated cell sorter, and infected with luciferase-reporter HIV-1 pseudotyped with CCR5-using Envs including R5 and R5X4. R5 and dual-R5 HIV-1 substantially infected BiFC-negative fraction rather than BiFC-positive fraction, indicating the preferential recognition of monomeric CCR5 by R5 and dual-R5 Envs. Although CCR5 antagonists enhanced oligomerization of CCR5, MVC-resistant HIV-1 was found to still recognize both MVC-bound and -unbound forms of monomeric CCR5, suggesting the constrained use of monomeric CCR5 by R5 HIV-1.