Analysis of HIV-1CRF_01 A/E protease inhibitor resistance: Structural determinants for maintaining sensitivity and developing resistance to atazanavir
Analysis of HIV-1CRF_01 A/E protease inhibitor resistance: Structural determinants for maintaining sensitivity and developing resistance to atazanavir
复制标题
DOI：
10.1021/bi051886s
复制
发表时间：
2006-05-02
期刊：
影响因子：
2.9
通讯作者：
Dunn, BM
中科院分区：
文献类型：
--
作者：
Clemente, JC;Coman, RM;Dunn, BM
为了分析HIV-1CRF_01A/E(AE)蛋白水解酶与B(B)亚型蛋白水解酶的天然多态和治疗选择性(活性和非活性部位)突变对耐药性的影响，我们设计了一系列HIV-1蛋白水解酶突变体。使用显色底物对这些变异体进行动力学分析表明，治疗前B和AE蛋白水解酶的底物特异性不同。利托那韦、依地那韦、奈非那韦、氨丙那韦、沙奎那韦、洛比那韦和阿扎那韦的抑制率分析显示，A/E中发现的自然基因多态性可以影响抑制剂的耐药性。同样明显的是，与B酶一样，A/E酶的高水平抗性是由于它获得了活性位点和非活性位点突变的组合。对阿扎那韦与治疗前B蛋白水解酶结合的结构分析表明，阿扎那韦与含有某些耐药突变的变异体保持结合亲和力的能力是由于它与瓣膜残基的独特相互作用。这种结构也解释了为什么150L和184V突变在降低阿扎那韦的结合亲和力方面是重要的。
A series of HIV-1 protease mutants has been designed in an effort to analyze the contribution to drug resistance provided by natural polymorphisms as well as therapy-selective (active and non-active site) mutations in the HIV-1 CRF_01 A/E (AE) protease when compared to that of the subtype B (B) protease. Kinetic analysis of these variants using chromogenic substrates showed differences in substrate specificity between pretherapy B and AE proteases. Inhibition analysis with ritonavir, indinavir, nelfinavir, amprenavir, saquinavir, lopinavir, and atazanavir revealed that the natural polymorphisms found in A/E can influence inhibitor resistance. It was also apparent that a high level of resistance in the A/E protease, as with B protease, is due to it aquiring a combination of active site and non-active site mutations. Structural analysis of atazanavir bound to a pretherapy B protease showed that the ability of atazanavir to maintain its binding affinity for variants containing some resistance mutations is due to its unique interactions with flap residues. This structure also explains why the 150L and 184V mutations are important in decreasing the binding affinity of atazanavir.