Probing Structural Changes among Analogous Inhibitor-Bound Forms of HIV-1 Protease and a Drug-Resistant Mutant in Solution by Nuclear Magnetic Resonance.
Probing Structural Changes among Analogous Inhibitor-Bound Forms of HIV-1 Protease and a Drug-Resistant Mutant in Solution by Nuclear Magnetic Resonance.
复制标题
通过核磁共振探测溶液中类似抑制剂结合形式的 HIV-1 蛋白酶和耐药突变体之间的结构变化。
DOI：
10.1021/acs.biochem.7b01238
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发表时间：
2018
期刊：
影响因子：
2.9
通讯作者：
Ishima,Rieko
中科院分区：
文献类型：
--
作者：
Khan,ShahidN;Persons,JohnD;Paulsen,JanetL;Guerrero,Michel;Schiffer,CeliaA;Kurt-Yilmaz,Nese;Ishima,Rieko
在最先进的抑制剂设计和高分辨率结构研究的时代，检测到的重要，但小的蛋白质结构差异，在走廊结合的形式是至关重要的，以进一步开发抑制剂。在这里，我们探讨了HIV-1蛋白酶（PR）构象之间的差异，地瑞那韦和四个类似的结合形式，并比较它们与耐药突变体使用核磁共振化学位移。这些受体结合形式中野生型（WT）PR的酰胺化学位移变化ΔCSP是微妙的，但可检测到，并且从受体结合位点延伸>10 μ m，在PR的两个亚基之间不对称。分子动力学模拟显示，差异局部氢键是这种远程不对称变化的分子基础。耐药突变体的抑制剂结合形式也显示出类似的长程ΔCSP模式。WT和突变体的ΔCSP值（ΔΔ CSP）的差异在寡核苷酸结合位点和周围区域观察到。比较高度相似的抑制剂和ΔΔ CSP之间的化学位移变化，有效地消除了不同化学基团引起的局部环境效应，并使这些敏感参数能够用于检测细微的蛋白质构象变化，并阐明抑制剂相互作用后的不对称和远程构象效应。
In the era of state-of-the-art inhibitor design and high-resolution structural studies, detection of significant but small protein structural differences in the inhibitor-bound forms is critical to further developing the inhibitor. Here, we probed differences in HIV-1 protease (PR) conformation among darunavir and four analogous inhibitor-bound forms and compared them with a drug-resistant mutant using nuclear magnetic resonance chemical shifts. Changes in amide chemical shifts of wild-type (WT) PR among these inhibitor-bound forms, ΔCSP, were subtle but detectable and extended >10 Å from the inhibitor-binding site, asymmetrically between the two subunits of PR. Molecular dynamics simulations revealed differential local hydrogen bonding as the molecular basis of this remote asymmetric change. Inhibitor-bound forms of the drug-resistant mutant also showed a similar long-range ΔCSP pattern. Differences in ΔCSP values of the WT and the mutant (ΔΔCSPs) were observed at the inhibitor-binding site and in the surrounding region. Comparing chemical shift changes among highly analogous inhibitors and ΔΔCSPs effectively eliminated local environmental effects stemming from different chemical groups and enabled exploitation of these sensitive parameters to detect subtle protein conformational changes and to elucidate asymmetric and remote conformational effects upon inhibitor interaction.