Recognition and blocking of HIV-1 gp41 pre-transmembrane sequence by monoclonal 4E10 antibody in a raft-like membrane environment
Recognition and blocking of HIV-1 gp41 pre-transmembrane sequence by monoclonal 4E10 antibody in a raft-like membrane environment
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DOI：
10.1074/jbc.m605998200
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发表时间：
2006-12-22
影响因子：
4.8
通讯作者：
Nieva, Jose L.
中科院分区：
文献类型：
--
作者：
Lorizate, Maier;Cruz, Antonio;Nieva, Jose L.
人免疫缺陷病毒（HIV-1）gp 41糖蛋白亚基跨膜锚之前的保守（664）DKWASLWNWFNITNWLWYIK（68）3（preTM）序列可被广泛中和的4 E10抗体接近，因此构成疫苗设计的潜在靶标。最近报道的结构数据与preTM插入到gp 41融合前状态的病毒外膜单层中是一致的（Zhu，P.，刘杰，贝丝，J.，Chertova，E.，Lifson，J.D.，格里斯，H.，奥费克湾一、Taylor，K.一、Roux，K. H.（2006）Nature 441，847-852）。在这里，我们证明了广泛中和的4 E10抗体能够特异性地阻断插入膜中的肽模拟物的膜重组活性。识别和重组阻断发生在胆固醇的存在下，而跨膜版本的1-棕榈酰-2-油酰磷脂酰胆碱：鞘磷脂混合物中促进不能有效地逮捕。荧光分光光度测定法使用罗丹明标记的肽显示，识别相关性更好地与孔形成阻断比与膜融合抑制。采用平面支持的脂质层和荧光显微镜进一步证实了抗体在筏状环境中识别前TM肽的能力。这些数据支持4 E10的膜结合表位识别导致前TM在膜界面处的聚类重组。我们建议，这一过程可能会干扰融合能力的复合物的形成在低尖峰密度存在于HIV-1膜。这项工作包括第一个实验报告的脂质调节抗体的能力，结合膜结合表位序列，并逮捕其重组活动。
The conserved (664)DKWASLWNWFNITNWLWYIK(68)3 (preTM) sequence preceding the transmembrane anchor of human immunodeficiency virus (HIV-1) gp41 glycoprotein subunit is accessible to the broadly neutralizing 4E10 antibody and, therefore, constitutes a potential target for vaccine design. Recently reported structural data are compatible with preTM insertion into the viral external membrane monolayer in the gp41 pre-fusion state (Zhu, P., Liu, J., Bess, J., Chertova, E., Lifson, J. D., Grise, H., Ofek, G. A., Taylor, K. A., and Roux, K. H. ( 2006) Nature 441, 847-852). Here we demonstrate that the broadly neutralizing 4E10 antibody is able to specifically block the membrane-restructuring activity of a peptide mimic inserted into membranes. Recognition and restructuring blocking occurred in the presence of cholesterol, whereas transmembrane versions as those promoted in 1-palmitoyl-2- oleoylphosphatidylcholine: sphingomyelin mixtures could not be effectively arrested. Spectrofluorimetric assays using rhodamine- labeled peptides revealed that recognition correlated better with pore-formation blocking than with membrane-fusion inhibition. The capacity of the antibody to recognize preTM peptides in a raft-like environment was further corroborated employing planar-supported lipid layers and fluorescence microscopy. These data support that membrane-bound epitope recognition by 4E10 results in clustering reorganization of preTM at the membrane interface. We propose that this process might interfere with the formation of fusion-competent complexes at the low spike densities existing in the HIV-1 membrane. This work comprises the first experimental report on a lipid-modulated antibody capacity to bind a membrane-bound epitope sequence and arrest its restructuring activity.