دورية أكاديمية
Small-Molecule Fusion Inhibitors Bind the pH-Sensing Stable Signal Peptide-GP2 Subunit Interface of the Lassa Virus Envelope Glycoprotein.
| العنوان: | Small-Molecule Fusion Inhibitors Bind the pH-Sensing Stable Signal Peptide-GP2 Subunit Interface of the Lassa Virus Envelope Glycoprotein. |
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| المؤلفون: | Shankar S; Montana Biotechnology Center, The University of Montana, Missoula, Montana, USA Division of Biological Sciences, The University of Montana, Missoula, Montana, USA., Whitby LR; Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA., Casquilho-Gray HE; Montana Biotechnology Center, The University of Montana, Missoula, Montana, USA., York J; Montana Biotechnology Center, The University of Montana, Missoula, Montana, USA., Boger DL; Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA., Nunberg JH; Montana Biotechnology Center, The University of Montana, Missoula, Montana, USA jack.nunberg@umontana.edu. |
| المصدر: | Journal of virology [J Virol] 2016 Jul 11; Vol. 90 (15), pp. 6799-807. Date of Electronic Publication: 2016 Jul 11 (Print Publication: 2016). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society For Microbiology Country of Publication: United States NLM ID: 0113724 Publication Model: Electronic-Print Cited Medium: Internet ISSN: 1098-5514 (Electronic) Linking ISSN: 0022538X NLM ISO Abbreviation: J Virol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Washington Dc : American Society For Microbiology Original Publication: Baltimore, American Society for Microbiology. |
| مواضيع طبية MeSH: | Protein Sorting Signals*, Antiviral Agents/*pharmacology , Membrane Fusion/*drug effects , Small Molecule Libraries/*pharmacology , Viral Envelope Proteins/*metabolism , Virus Internalization/*drug effects, Animals ; Chlorocebus aethiops ; Humans ; Hydrogen-Ion Concentration ; Lassa virus ; Protein Subunits ; Vero Cells ; Viral Envelope Proteins/chemistry |
| مستخلص: | Unlabelled: Arenavirus species are responsible for severe life-threatening hemorrhagic fevers in western Africa and South America. Without effective antiviral therapies or vaccines, these viruses pose serious public health and biodefense concerns. Chemically distinct small-molecule inhibitors of arenavirus entry have recently been identified and shown to act on the arenavirus envelope glycoprotein (GPC) to prevent membrane fusion. In the tripartite GPC complex, pH-dependent membrane fusion is triggered through a poorly understood interaction between the stable signal peptide (SSP) and the transmembrane fusion subunit GP2, and our genetic studies have suggested that these small-molecule inhibitors act at this interface to antagonize fusion activation. Here, we have designed and synthesized photoaffinity derivatives of the 4-acyl-1,6-dialkylpiperazin-2-one class of fusion inhibitors and demonstrate specific labeling of both the SSP and GP2 subunits in a native-like Lassa virus (LASV) GPC trimer expressed in insect cells. Photoaddition is competed by the parental inhibitor and other chemically distinct compounds active against LASV, but not those specific to New World arenaviruses. These studies provide direct physical evidence that these inhibitors bind at the SSP-GP2 interface. We also find that GPC containing the uncleaved GP1-GP2 precursor is not susceptible to photo-cross-linking, suggesting that proteolytic maturation is accompanied by conformational changes at this site. Detailed mapping of residues modified by the photoaffinity adducts may provide insight to guide the further development of these promising lead compounds as potential therapeutic agents to treat Lassa hemorrhagic fever. Importance: Hemorrhagic fever arenaviruses cause lethal infections in humans and, in the absence of licensed vaccines or specific antiviral therapies, are recognized to pose significant threats to public health and biodefense. Lead small-molecule inhibitors that target the arenavirus envelope glycoprotein (GPC) have recently been identified and shown to block GPC-mediated fusion of the viral and cellular endosomal membranes, thereby preventing virus entry into the host cell. Genetic studies suggest that these inhibitors act through a unique pH-sensing intersubunit interface in GPC, but atomic-level structural information is unavailable. In this report, we utilize novel photoreactive fusion inhibitors and photoaffinity labeling to obtain direct physical evidence for inhibitor binding at this critical interface in Lassa virus GPC. Future identification of modified residues at the inhibitor-binding site will help elucidate the molecular basis for fusion activation and its inhibition and guide the development of effective therapies to treat arenaviral hemorrhagic fevers. (Copyright © 2016, American Society for Microbiology. All Rights Reserved.) |
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| معلومات مُعتمدة: | R21 AI120490 United States AI NIAID NIH HHS; R01 CA042056 United States CA NCI NIH HHS; R01 AI074818 United States AI NIAID NIH HHS; P20 GM103546 United States GM NIGMS NIH HHS; U54 AI065357 United States AI NIAID NIH HHS |
| المشرفين على المادة: | 0 (Antiviral Agents) 0 (Protein Sorting Signals) 0 (Protein Subunits) 0 (Small Molecule Libraries) 0 (Viral Envelope Proteins) |
| تواريخ الأحداث: | Date Created: 20160520 Date Completed: 20170602 Latest Revision: 20200401 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC4944282 |
| DOI: | 10.1128/JVI.00597-16 |
| PMID: | 27194767 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1098-5514 |
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| DOI: | 10.1128/JVI.00597-16 |