The connection domain is implicated in metalloporphyrin binding and inhibition of HIV reverse transcriptase
The connection domain is implicated in metalloporphyrin binding and inhibition of HIV reverse transcriptase
复制标题
DOI：
10.1074/jbc.274.3.1549
复制
发表时间：
1999-01-15
影响因子：
4.8
通讯作者：
Paterson, Y
中科院分区：
文献类型：
--
作者：
Argyris, EG;Vanderkooi, JM;Paterson, Y
我们已经证明，血红素和锌原卟啉对人类免疫缺陷病毒1型(HIV-1)和2型(HIV-2)逆转录酶(RTS)都有抑制作用，并与其他核苷和非核苷抑制剂一起对HIV-1RT抑制起相加作用。从HIV-1和HIV-2 RT的连接亚区中分离到一个序列与398-407序列相似的抗血红素噬菌体多肽库，这表明HIV RT的这个高度保守的区域对应于金属卟啉的结合部位，而与HIV-1 RT的398-407序列相对应的合成肽在RT抑制实验中对金属卟啉的抑制具有保护作用，因为它能够逆转这两种金属卟啉对HIV-1 RT活性的抑制作用。此外，本征荧光分析表明，这些金属卟啉与合成肽398-407以及完整的二聚体HIV-1RT结合。这个新的抑制位点的发现将有助于扩大我们对金属卟啉在RT抑制中的作用轨迹的理解，并将有助于设计和开发更有效的金属卟啉RT抑制剂来管理HIV感染。
We have shown that heme and zinc protoporphyrin inhibit both human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) reverse transcriptases (RTs) and, in combination with other nucleoside and non-nucleoside inhibitors, exert an additive effect on HIV-1 RT inhibition. Screening of a phage peptide library against heme resulted in the isolation of a peptide with sequence similarity to sequence 398-407 from the connection subdomain of both HIV-1 and HIV-2 RTs, suggesting that this highly conserved region of HIV RTs corresponds to the binding site for metalloporphyrins and a new site for inhibition of enzyme activity Inclusion of a synthetic peptide corresponding to the exact sequence 398-407 of HIV-1 RT in RT inhibition assays had a protective effect on metalloporphyrin inhibition, as it was able to reverse the inhibitory effect of both metalloporphyrins on HIV-1 RT activity. Furthermore, intrinsic fluorescence assays indicated that these metalloporphyrins bind to synthetic peptide 398-407 as well as to intact dimeric HIV-1 RT. The identification of this novel inhibition site will help to expand our understanding of the rode of action of metalloporphyrins in RT inhibition and will assist in the design and development of more potent metalloporphyrin RT inhibitors for the management of HIV infection.