Peptides derived from the reverse transcriptase of human immunodeficiency virus type 1 as novel inhibitors of the viral integrase
Peptides derived from the reverse transcriptase of human immunodeficiency virus type 1 as novel inhibitors of the viral integrase
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DOI：
10.1074/jbc.m414679200
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发表时间：
2005-06-10
影响因子：
4.8
通讯作者：
Hizi, A
中科院分区：
文献类型：
--
作者：
Gleenberg, IO;Avidan, O;Hizi, A
最近的研究表明，HIV-1的整合酶（IN）在体外被HIV-1逆转录酶（RT）抑制。我们进一步研究了特定的蛋白质序列的RT参与这种抑制筛选一个完整的库的RT衍生肽抑制IN活动。两个20个残基的肽，肽4286，来自RT DNA聚合酶结构域，和一个指定的4321，从RT核糖核酸酶H结构域，在体外抑制IN的酶活性。前一种肽抑制所有三种IN相关活性（3 '端加工、链转移和分解），而后一种肽主要抑制前两种功能。我们显示了序列和肽长度对于有效抑制IN活性的重要性。肽与IN的结合测定（不存在DNA底物）表明两种抑制肽（以及几种非抑制肽）直接与IN相互作用。此外，分离的IN催化核心结构域也直接与两个抑制肽相互作用。然而，只有肽4286可以抑制与IN核心结构域相关的崩解活性，因为这种活性是该结构域表现出的唯一活性。该结果是由于肽4321对全长IN的崩解缺乏抑制而预期的。所提出的数据和三维模型表明，这种抑制作用是由于IN催化结构域的空间位阻所致。这些信息可以大大促进针对HIV INs的新药的开发，从而有助于对抗艾滋病。
Recent studies have shown that the integrase ( IN) of HIV-1 is inhibited in vitro by HIV-1 reverse transcriptase (RT). We further investigated the specific protein sequences of RT that were involved in this inhibition by screening a complete library of RT-derived peptides for their inhibition of IN activities. Two 20-residue peptides, peptide 4286, derived from the RT DNA polymerase domain, and the one designated 4321, from the RT ribonuclease H domain, inhibit the enzymatic activities of IN in vitro. The former peptide inhibits all three IN-associated activities (3'-end processing, strand transfer, and disintegration), whereas the latter one inhibits primarily the first two functions. We showed the importance of the sequences and peptide length for the effective inhibition of IN activities. Binding assays of the peptides to IN ( with no DNA substrate present) indicated that the two inhibitory peptides ( as well as several non-inhibitory peptides) interact directly with IN. Moreover, the isolated catalytic core domain of IN also interacted directly with the two inhibitory peptides. Nevertheless, only peptide 4286 can inhibit the disintegration activity associated with the IN core domain, because this activity is the only one exhibited by this domain. This result was expected from the lack of inhibition of disintegration of full-length IN by peptide 4321. The data and the three-dimensional models presented suggested that the inhibition resulted from steric hindrance of the catalytic domain of IN. This information can substantially facilitate the development of novel drugs against HIV INs and thus contribute to the fight against AIDS.