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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Gao, Guofen Peachman, Kristina K. Polonis, Victoria R. Rao, Mangala Alving, Carl R. Rao, Venigalla B. Wieczorek, Lindsay |
| Description | Author Affiliation: Gao G ( Department of Biology, The Catholic University of America, Washington, DC 20064, USA.) |
| Abstract | The HIV-1 envelope spike is a trimer of heterodimers composed of an external glycoprotein gp120 and a transmembrane glycoprotein gp41. gp120 initiates virus entry by binding to host receptors, whereas gp41 mediates fusion between viral and host membranes. Although the basic pathway of HIV-1 entry has been extensively studied, the detailed mechanism is still poorly understood. Design of gp41 recombinants that mimic key intermediates is essential to elucidate the mechanism as well as to develop potent therapeutics and vaccines. Here, using molecular genetics and biochemical approaches, a series of hypotheses was tested to overcome the extreme hydrophobicity of HIV-1 gp41 and design a soluble near full-length gp41 trimer. The two long heptad repeat helices HR1 and HR2 of gp41 ectodomain were mutated to disrupt intramolecular HR1-HR2 interactions but not intermolecular HR1-HR1 interactions. This resulted in reduced aggregation and improved solubility. Attachment of a 27-amino acid foldon at the C terminus and slow refolding channeled gp41 into trimers. The trimers appear to be stabilized in a prehairpin-like structure, as evident from binding of a HR2 peptide to exposed HR1 grooves, lack of binding to hexa-helical bundle-specific NC-1 mAb, and inhibition of virus neutralization by broadly neutralizing antibodies 2F5 and 4E10. Fusion to T4 small outer capsid protein, Soc, allowed display of gp41 trimers on the phage nanoparticle. These approaches for the first time led to the design of a soluble gp41 trimer containing both the fusion peptide and the cytoplasmic domain, providing insights into the mechanism of entry and development of gp41-based HIV-1 vaccines. |
| ISSN | 00219258 |
| e-ISSN | 1083351X |
| Journal | Journal of Biological Chemistry |
| Issue Number | 1 |
| Volume Number | 288 |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date | 2013-01-04 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Anti-HIV Agents Pharmacology Antibodies, Monoclonal Chemistry HIV Envelope Protein Gp41 HIV-1 Metabolism Biochemistry Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Epitopes Genetic Vectors HIV Fusion Inhibitors Mutagenesis Mutation Peptide Library Peptides Protein Conformation Protein Folding Protein Structure, Tertiary Recombinant Proteins Virus Internalization Drug Effects Research Support, N.I.H., Extramural Molecular Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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