Dilation of the human immunodeficiency virus-1 envelope glycoprotein fusion pore revealed by the inhibitory action of a synthetic peptide from gp41.
Dilation of the human immunodeficiency virus-1 envelope glycoprotein fusion pore revealed by the inhibitory action of a synthetic peptide from gp41.
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DOI：
10.1083/jcb.140.2.315
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发表时间：
1998-01-26
期刊：
影响因子：
--
通讯作者：
Blumenthal R
中科院分区：
文献类型：
--
作者：
Muñoz-Barroso I;Durell S;Sakaguchi K;Appella E;Blumenthal R
我们监测了由单个人类免疫缺陷病毒(HIV-1)包膜糖蛋白表达细胞和CD4+靶细胞组成的细胞对之间的融合，该靶细胞已经被膜上的荧光脂和胞浆中的荧光溶质标记。我们开发了一种新的三色检测方法，以跟踪荧光脂类和/或溶质重新分布到细胞中的情况。脂类和溶质重分布分别是由于打开允许脂质的融合孔和允许溶质的融合孔(FPS)而发生的。一种合成肽(DP178)，对应于HIV-1LAI gp120-gp41序列的643-678残基(Wild，C.T.，D.C.Shugars，T.K.Greenwell，C.B.McDanal和T.J.Matthews。1994年。程序娜塔莉。阿卡德。SCI。美国。91：12676-12680)在50 ng/ml时完全抑制FPS，而在该浓度下有20-30%的融合活性(通过脂质重分布来衡量)。表达gp120-41的细胞与靶细胞共培养6h后，在脂质混合与内容物混合方面的差异一直保持到6h，这表明DP178可以在很长一段时间内“夹住”脂质混合中间体中的融合复合体。来自gp41 NH2末端的一个肽DP107在较高浓度下抑制HIV-1LAI gp120-gp41介导的细胞融合，但在不同的抑制剂浓度下，脂类和水中染料的再分布没有差异。DP178对溶质重分布的抑制作用在共培养的前15分钟内完全被加入融合反应混合物中。我们根据融合的孔扩张模型分析了抑制数据，该模型结合了最近确定的gp41核心的高分辨率结构。
We have monitored fusion between cell pairs consisting of a single human immunodeficiency virus–1 (HIV-1) envelope glycoprotein–expressing cell and a CD4+ target cell, which had been labeled with both a fluorescent lipid in the membrane and a fluorescent solute in the cytosol. We developed a new three-color assay to keep track of the cell into which fluorescent lipids and/or solutes are redistributed. Lipid and solute redistribution occur as a result of opening a lipid-permissive fusion pore and a solute-permissive fusion pore (FPS), respectively. A synthetic peptide (DP178) corresponding to residues 643–678 of the HIV-1LAI gp120-gp41 sequence (Wild, C.T., D.C. Shugars, T.K. Greenwell, C.B. McDanal, and T.J. Matthews. 1994. Proc. Natl. Acad. Sci. USA. 91:12676–12680) completely inhibited FPS at 50 ng/ml, whereas at that concentration there was 20–30% fusion activity measured by the lipid redistribution. The differences detected in lipid mixing versus contents mixing are maintained up to 6 h of coculture of gp120-41–expressing cells with target cells, indicating that DP178 can “clamp” the fusion complex in the lipid mixing intermediate for very long time periods. A peptide from the NH2-terminal of gp41, DP107, inhibited HIV-1LAI gp120-gp41–mediated cell fusion at higher concentrations, but with no differences between lipid and aqueous dye redistribution at the different inhibitor concentrations. The inhibition of solute redistribution by DP178 was complete when the peptide was added to the fusion reaction mixture during the first 15 min of coculture. We have analyzed the inhibition data in terms of a fusion pore dilation model that incorporates the recently determined high resolution structure of the gp41 core.
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DOI：
10.1083/jcb.131.3.679
发表时间：
1995-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Melikyan GB;White JM;Cohen FS
通讯作者：
Cohen FS
影响因子：
56.9
作者：
Feng, Y;Broder, CC;Berger, EA
通讯作者：
Berger, EA
影响因子：
5.4
作者：
CHEN, CH;MATTHEWS, TJ;GREENBERG, ML
通讯作者：
GREENBERG, ML
影响因子：
2.9
作者：
ELLENS, H;BENTZ, J;WHITE, JM
通讯作者：
WHITE, JM
影响因子：
5.4
作者：
FU, YK;HART, TK;BUGELSKI, PJ
通讯作者：
BUGELSKI, PJ