The Anti-HIV pentameric pseudopeptide HB-19 binds the C-terminal end of nucleolin and prevents anchorage of virus particles in the plasma membrane of target cells
The Anti-HIV pentameric pseudopeptide HB-19 binds the C-terminal end of nucleolin and prevents anchorage of virus particles in the plasma membrane of target cells
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DOI：
10.1074/jbc.m110024200
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发表时间：
2002-06-07
影响因子：
4.8
通讯作者：
Hovanessian, AG
中科院分区：
文献类型：
--
作者：
Nisole, S;Said, EA;Hovanessian, AG
多价伪肽 HB-19 与细胞表面表达的核仁素结合，通过阻止病毒颗粒附着并因此锚定在质膜上，成为人类免疫缺陷病毒 (HIV) 感染的有效抑制剂。我们表明，表面结合的 HB-19A（类似于 HB-19，但具有修改的模板）的交联导致 HB-19A 与表面核仁素聚集。与其特定作用一致，HB-19A 与不同类型细胞的结合达到饱和浓度，据报道可抑制 HrV 感染。通过使用中国仓鼠卵巢突变细胞系，我们证实IFM-19A与表面核仁素的结合不依赖于乙酰肝素和硫酸软骨素蛋白聚糖。体外生成的全长核仁素被发现可以结合 HB-19A，而含有核仁素酸性氨基酸片段的 N 末端部分则不能。使用核仁素 C 端部分的各种缺失构建体可以鉴定核仁素 C 端末端，其中包含氨基酸基序 RGG 的重复序列，作为结合 HB-19A 的结构域。最后，对应于最后 C 端 63 个氨基酸的合成肽能够在 HIV 附着到细胞的阶段抑制 HIV 感染，从而表明该结构域可能在 HIV 锚定过程中发挥作用。
The multivalent pseudopeptide HB-19 that binds the cell-surface-expressed nucleolin is a potent inhibitor of human immunodeficiency virus (HIV) infection by blocking virus particle attachment and thus anchorage in the plasma membrane. We show that cross-linking of surface-bound HB-19A (like HB-19 but with a modified template) results in aggregation of HB-19A with surface nucleolin. Consistent with its specific action, HB-19A binding to different types of cells reaches saturation at concentrations that have been reported to result in inhibition of HrV infection. By using Chinese hamster ovary mutant cell lines, we confirm that the binding of IFM-19A to surface nucleolin is independent of heparan and chondroitin sulfate proteoglyeans. In vitro generated full-length nucleolin was found to bind HB-19A, whereas the N-terminal part containing the acidic amino acid stretches of nucleolin did not. The use of various deletion constructs of the C-terminal part of nucleolin then permitted the identification of the extreme C-terminal end of nucleolin, containing repeats of the amino acid motif, RGG, as the domain that binds HB-19A. Finally, a synthetic peptide corresponding to the last C-terminal 63 amino acids was able to inhibit HIV infection at the stage of HIV attachment to cells, thus suggesting that this domain could be functional in the HIV anchorage process.