Monoclonal antibodies against human immunodeficiency virus type 1 integrase: epitope mapping and differential effects on integrase activities in vitro.
Monoclonal antibodies against human immunodeficiency virus type 1 integrase: epitope mapping and differential effects on integrase activities in vitro.
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抗人类免疫缺陷病毒 1 型整合酶的单克隆抗体：表位作图和体外对整合酶活性的差异影响。
DOI：
10.1128/jvi.70.3.1580-1587.1996
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Helland,DE
中科院分区：
文献类型：
--
作者：
Nilsen,BM;Haugan,IR;Berg,K;Olsen,L;Brown,PO;Helland,DE
人类免疫缺陷病毒1型（HIV-1）整合酶（IN）催化病毒DNA整合到宿主染色体上，这是逆转录病毒复制的重要步骤。作为研究该酶结构和功能的工具，制备了针对HIV-1 IN的单克隆抗体（mab）。表位定位表明，获得的17个单克隆抗体可分为7个不同的组，并对代表这些组的单克隆抗体进行了筛选，以检测其对体外in活性的影响。四组单克隆抗体识别的表位位于in的N端附近保守的HHCC基序内和周围的氨基酸（aa） 1 ~ 16、17 ~ 38或42 ~ 55区域。与这些表位结合的单克隆抗体抑制末端加工和DNA连接，刺激或几乎不影响IN的解体和重新整合活性。两个单抗结合在蛋白中心核心的aa 56 ~ 102区域或c端一半的aa 186 ~ 250区域，对in的体外活性影响较小。在HIV-1 IN的aa262 ~ 271区表位上识别的3个单抗与HIV-2 IN发生交叉反应。与该表位结合的单克隆抗体明显抑制末端加工和DNA连接，并刺激或几乎不影响崩解。与n端特异性单克隆抗体相比，这些c端特异性单克隆抗体消除了In的重新整合活性。
Human immunodeficiency virus type 1 (HIV-1) integrase (IN) catalyzes the integration of viral DNA into the host chromosome, an essential step in retroviral replication. As a tool to study the structure and function of this enzyme, monoclonal antibodies (MAbs) against HIV-1 IN were produced. Epitope mapping demonstrated that the 17 MAbs obtained could be divided into seven different groups, and the selection of MAbs representing these groups were tested for their effect on in vitro activities of IN. Four groups of MAbs recognized epitopes within the region of amino acids (aa) 1 to 16, 17 to 38, or 42 to 55 in and around the conserved HHCC motif near the N terminus of IN. MAbs binding to these epitopes inhibited end processing and DNA joining and either stimulated or had little effect on disintegration and reintegration activities of IN. Two MAbs binding to epitopes within the region of aa 56 to 102 in the central core or aa 186 to 250 in the C-terminal half of the protein showed only minor effects on the in vitro activities of IN. Three Mabs which recognized on epitope within the region of aa262 to 271 of HIV-1 IN cross-reacted with HIV-2 IN. MAbs binding to this epitope clearly inhibited end processing and DNA joining and stimulated or had little effect on disintegration. In contrast to the N-terminal-specific MAbs, these C-terminal-specific MAbs abolished reintegration activity of IN.