Structure of a covalently trapped catalytic complex of HIV-I reverse transcriptase: Implications for drug resistance
Structure of a covalently trapped catalytic complex of HIV-I reverse transcriptase: Implications for drug resistance
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DOI：
10.1126/science.282.5394.1669
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发表时间：
1998-11-27
期刊：
影响因子：
56.9
通讯作者：
Harrison, SC
中科院分区：
文献类型：
--
作者：
Huang, HF;Chopra, R;Harrison, SC
以DNA为模板，用二硫键结合三磷酸脱氧核苷(DNTP)制备了人类免疫缺陷病毒1型(HIV-1)逆转录酶停滞复合体，测定了该复合体的晶体结构。在3‘-引物末端的双脱氧核苷酸的存在允许捕获底物准备攻击dNTP的状态。伴随催化复合体形成的构象变化会产生不同的残基簇，这些残基在对核苷类似药物具有耐药性的病毒中发生改变。这些残基在dNTP附近的定位有助于解决一些长期存在的关于抗性分子基础的谜题。相对于正常的dNTPs，耐药突变可能会影响抑制剂的结合或反应活性，并且突变的聚集与药物的化学结构有关。
A combinatorial disulfide cross-linking strategy was used to prepare a stalled complex of human immunodeficiency virus-type 1 (HIV-1) reverse transcriptase with a DNA template:primer and a deoxynucleoside triphosphate (dNTP), and the crystal structure of the complex was determined at a resolution of 3.2 angstroms. The presence of a dideoxynucleotide at the 3'-primer terminus allows capture of a state in which the substrates are poised for attack on the dNTP. Conformational changes that accompany formation of the catalytic complex produce distinct clusters of the residues that are altered in viruses resistant to nucleoside analog drugs. The positioning of these residues in the neighborhood of the dNTP helps to resolve some Long-standing puzzles about the molecular basis of resistance. The resistance mutations are Likely to influence binding or reactivity of the inhibitors, relative to normal dNTPs, and the clustering of the mutations correlates with the chemical structure of the drug.