Secondary structure and membrane topology of dengue virus NS4B N-terminal 125 amino acids.
| Autor: | Li Y; Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore., Kim YM; Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore., Zou J; Novartis Institute for Tropical Diseases, Singapore, Singapore., Wang QY; Novartis Institute for Tropical Diseases, Singapore, Singapore., Gayen S; Department of Pharmaceutical Sciences, Dr H. S. Gour University, Sagar 470003, Madhya Pradesh, India., Wong YL; Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore., Lee le T; Novartis Institute for Tropical Diseases, Singapore, Singapore., Xie X; Novartis Institute for Tropical Diseases, Singapore, Singapore., Huang Q; Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore., Lescar J; School of Biological Sciences, Nanyang Technological University, Singapore, Singapore., Shi PY; Novartis Institute for Tropical Diseases, Singapore, Singapore. Electronic address: peshi@utmb.edu., Kang C; Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore. Electronic address: cbkang@etc.a-star.edu.sg. |
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| Jazyk: | angličtina |
| Zdroj: | Biochimica et biophysica acta [Biochim Biophys Acta] 2015 Dec; Vol. 1848 (12), pp. 3150-7. Date of Electronic Publication: 2015 Sep 25. |
| DOI: | 10.1016/j.bbamem.2015.09.016 |
| Abstrakt: | The transmembrane NS4B protein of dengue virus (DENV) is a validated antiviral target that plays important roles in viral replication and invasion of innate immune response. The first 125 amino acids of DENV NS4B are sufficient for inhibition of alpha/beta interferon signaling. Resistance mutations to NS4B inhibitors are all mapped to the first 125 amino acids. In this study, we expressed and purified a protein representing the first 125 amino acids of NS4B (NS4B(1-125)). This recombinant NS4B(1-125) protein was reconstituted into detergent micelles. Solution NMR spectroscopy demonstrated that there are five helices (α1 to α5) present in NS4B(1-125). Dynamic studies, together with a paramagnetic relaxation enhancement experiment demonstrated that four helices, α2, α3, α4, and α5 are embedded in the detergent micelles. Comparison of wild type and V63I mutant (a mutation that confers resistance to NS4B inhibitor) NS4B(1-125) proteins demonstrated that V63I mutation did not cause significant conformational changes, however, V63 may have a molecular interaction with residues in the α5 transmembrane domain under certain conditions. The structural and dynamic information obtained in study is helpful to understand the structure and function of NS4B. (Copyright © 2015 Elsevier B.V. All rights reserved.) |
| Databáze: | MEDLINE |
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