Journal article
Lytic Inactivation of Human Immunodeficiency Virus by Dual Engagement of gp120 and gp41 Domains in the Virus Env Protein Trimer
Biochemistry (Easton), v 55(44), pp 6100-6114
08 Nov 2016
PMID: 27731975
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We recently reported the discovery of a recombinant chimera, denoted DAVEI (dual-acting virucidal entry inhibitor), which is able to selectively cause specific and potent lytic inactivation of both pseudotyped and fully infectious human immunodeficiency virus (HIV-1) virions. The chimera is composed of the lectin cyanovirin-N (CVN) fused to the 20-residue membrane-proximal external region (MPER) of HIV-1 gp41. Because the Env gp120-binding CVN domain on its own is not lytic, we sought here to determine how the MPER
domain is able to endow the chimera with virolytic activity. We used a protein engineering strategy to identify molecular determinants of MPER
that are important for function. Recombinant mutagenesis and truncation demonstrated that the MPER
domain could be significantly minimized without loss of function. The dependence of lysis on specific MPER sequences of DAVEI, determination of minimal linker length, and competition by a simplified MPER surrogate peptide suggested that the MPER domain of DAVEI interacts with the Env spike trimer, likely with the gp41 region. This conclusion was further supported by observations from binding of the biotinylated MPER surrogate peptide to Env protein expressed on cells, monoclonal antibody competition, a direct binding enzyme-linked immunosorbent assay on viruses with varying numbers of trimeric spikes on their surfaces, and comparison of maximal interdomain spacing in DAVEI to that in high-resolution structures of Env. The finding that MPER
in CVN-MPER linker sequences can be minimized without loss of virolytic function provides an improved experimental path for constructing size-minimized DAVEI chimeras and molecular tools for determining how simultaneous engagement of gp120 and gp41 by these chimeras can disrupt the metastable virus Env spike.
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Details
- Title
- Lytic Inactivation of Human Immunodeficiency Virus by Dual Engagement of gp120 and gp41 Domains in the Virus Env Protein Trimer
- Creators
- Bibek Parajuli - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United StatesKriti Acharya - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United StatesReina Yu - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United StatesBrendon Ngo - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United StatesAdel A Rashad - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United StatesCameron F Abrams - Department of Chemical and Biological Engineering, Drexel University , Philadelphia, Pennsylvania 19104, United StatesIrwin M Chaiken - Department of Biochemistry and Molecular Biology, Drexel University College of Medicine , Philadelphia, Pennsylvania 19102, United States
- Publication Details
- Biochemistry (Easton), v 55(44), pp 6100-6114
- Publisher
- American Chemical Society; Washington, DC
- Grant note
- P01 GM056550 / NIGMS NIH HHS R01 GM115249 / NIGMS NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology; Chemical and Biological Engineering
- Web of Science ID
- WOS:000387518900003
- Scopus ID
- 2-s2.0-84994751026
- Other Identifier
- 991014877978404721
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- Web of Science research areas
- Biochemistry & Molecular Biology