Nakamura Akiyoshi, Tamura Noriko, Yasutake Yoshiaki
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo, Hokkaido 062-8517, Japan.
Acta Crystallogr F Struct Biol Commun. 2015 Nov;71(Pt 11):1384-90. doi: 10.1107/S2053230X15017896. Epub 2015 Oct 23.
Hepatitis B virus polymerase (HBV Pol) is an important target for anti-HBV drug development; however, its low solubility and stability in vitro has hindered detailed structural studies. Certain nucleotide reverse transcriptase (RT) inhibitors (NRTIs) such as tenofovir and lamivudine can inhibit both HBV Pol and Human immunodeficiency virus 1 (HIV-1) RT, leading to speculation on structural and mechanistic analogies between the deoxynucleotide triphosphate (dNTP)-binding sites of these enzymes. The Q151M mutation in HIV-1 RT, located at the dNTP-binding site, confers resistance to various NRTIs, while maintaining sensitivity to tenofovir and lamivudine. The residue corresponding to Gln151 is strictly conserved as a methionine in HBV Pol. Therefore, the structure of the dNTP-binding pocket of the HIV-1 RT Q151M mutant may reflect that of HBV Pol. Here, the crystal structure of HIV-1 RT Q151M, determined at 2.6 Å resolution, in a new crystal form with space group P321 is presented. Although the structure of HIV-1 RT Q151M superimposes well onto that of HIV-1 RT in a closed conformation, a slight movement of the β-strands (β2-β3) that partially create the dNTP-binding pocket was observed. This movement might be caused by the introduction of the bulky thioether group of Met151. The structure also highlighted the possibility that the hydrogen-bonding network among amino acids and NRTIs is rearranged by the Q151M mutation, leading to a difference in the affinity of NRTIs for HIV-1 RT and HBV Pol.
乙型肝炎病毒聚合酶(HBV Pol)是抗HBV药物研发的重要靶点;然而,其在体外的低溶解度和稳定性阻碍了详细的结构研究。某些核苷酸逆转录酶(RT)抑制剂,如替诺福韦和拉米夫定,既能抑制HBV Pol,也能抑制人类免疫缺陷病毒1型(HIV-1)RT,这引发了人们对这些酶的脱氧核苷酸三磷酸(dNTP)结合位点之间结构和机制相似性的猜测。位于dNTP结合位点的HIV-1 RT中的Q151M突变赋予了对各种NRTIs的抗性,同时保持对替诺福韦和拉米夫定的敏感性。在HBV Pol中,与Gln151对应的残基严格保守为甲硫氨酸。因此,HIV-1 RT Q151M突变体的dNTP结合口袋结构可能反映了HBV Pol的结构。在此,展示了以2.6 Å分辨率测定的HIV-1 RT Q151M在空间群为P321的新晶体形式中的晶体结构。尽管HIV-1 RT Q151M的结构与处于封闭构象的HIV-1 RT的结构很好地重叠,但观察到部分形成dNTP结合口袋的β链(β2-β3)有轻微移动。这种移动可能是由Met151的庞大硫醚基团的引入引起的。该结构还突出了这样一种可能性,即氨基酸与NRTIs之间的氢键网络因Q151M突变而重新排列,导致NRTIs对HIV-1 RT和HBV Pol的亲和力存在差异。