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从印度尼西亚食蟹猴中分离出的2型猿猴免疫缺陷病毒(SRV-2)逆转录酶初步三维结构的构建

Construction of A Preliminary Three-Dimensional Structure Serotype-2 (SRV-2) Reverse Transcriptase Isolated from Indonesian Cynomolgus Monkey.

作者信息

Saepuloh Uus, Iskandriati Diah, Pamungkas Joko, Solihin Dedy Duryadi, Mariya Sela Septima, Sajuthi Dondin

机构信息

Primate Research Centre, Bogor Agricultural University (PSSP LPPM IPB), Jalan Lodaya II/5 Bogor 16151, Indonesia.

Faculty of Veterinary Medicine, Bogor Agricultural University, Kampus Darmaga, Bogor 16680, Indonesia.

出版信息

Trop Life Sci Res. 2020 Oct;31(3):47-61. doi: 10.21315/tlsr2020.31.3.4. Epub 2020 Oct 15.

Abstract

serotype-2 (SRV-2) is an important pathogenic agent in Asian macaques. It is a potential confounding variable in biomedical research. SRV-2 also provides a valuable viral model compared to other retroviruses which can be used for understanding many aspects of retroviral-host interactions and immunosuppression, infection mechanism, retroviral structure, antiretroviral and vaccine development. In this study, we isolated the gene encoding reverse transcriptase enzyme (RT) of SRV-2 that infected Indonesian cynomolgus monkey (Mf ET1006) and predicted the three dimensional structure model using the iterative threading assembly refinement (I-TASSER) computational programme. This SRV-2 RT Mf ET1006 consisted of 547 amino acids at nucleotide position 3284-4925 of whole genome SRV-2. The polymerase active site located in the finger/palm subdomain characterised by three conserved catalytic aspartates (Asp90, Asp165, Asp166), and has a highly conserved YDD motif as Tyr163, Met164, Asp165 and Asp166. We estimated that this SRV-2 RT Mf ET1006 structure has the accuracy of template modelling score (TM-score 0.90 ± 0.06) and root mean square deviation (RMSD) 4.7 ± 3.1Å, indicating that this model can be trusted and the accuracy can be seen from the appearance of protein folding in tertiary structure. The superpositionings between SRV-2 RT Mf ET1006 and Human Immunodeficiency Virus-1 (HIV-1) RT were performed to predict the structural in details and to optimise the best fits for illustrations. This SRV-2 RT Mf ET1006 structure model has the highest homology to HIV-1 RT (2B6A.pdb) with estimated accuracy at TM-score 0.911, RMSD 1.85 Å, and coverage of 0.953. This preliminary study of SRV-2 RT Mf ET1006 structure modelling is intriguing and provide some information to explore the molecular characteristic and biochemical mechanism of this enzyme.

摘要

2型猿猴泡沫病毒(SRV-2)是亚洲猕猴中的一种重要病原体。它是生物医学研究中一个潜在的混杂变量。与其他逆转录病毒相比,SRV-2还提供了一个有价值的病毒模型,可用于理解逆转录病毒与宿主相互作用和免疫抑制、感染机制、逆转录病毒结构、抗逆转录病毒药物及疫苗开发等诸多方面。在本研究中,我们分离了感染印度尼西亚食蟹猴(Mf ET1006)的SRV-2编码逆转录酶(RT)的基因,并使用迭代穿线组装优化(I-TASSER)计算程序预测了三维结构模型。该SRV-2 RT Mf ET1006由全基因组SRV-2核苷酸位置3284-4925处的547个氨基酸组成。聚合酶活性位点位于指状/掌状亚结构域,其特征为三个保守的催化天冬氨酸(Asp90、Asp165、Asp166),并且具有高度保守的YDD基序,即Tyr163、Met164、Asp165和Asp166。我们估计该SRV-2 RT Mf ET1006结构的模板建模得分(TM-score 0.90±0.06)和均方根偏差(RMSD)为4.7±3.1Å,表明该模型可信,且从三级结构中蛋白质折叠的外观可看出其准确性。对SRV-2 RT Mf ET1006和人类免疫缺陷病毒1型(HIV-1)RT进行了叠加,以详细预测结构并优化插图的最佳匹配。该SRV-2 RT Mf ET1006结构模型与HIV-1 RT(2B6A.pdb)具有最高同源性,估计准确性为TM-score 0.911、RMSD 1.85 Å和覆盖率0.953。对SRV-2 RT Mf ET1006结构建模的这项初步研究很有趣,并为探索该酶的分子特征和生化机制提供了一些信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91e6/7652245/df4840df7c12/TLSR-31-3-47-g001.jpg

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