1. Physiological temperatures reduce dimerization of dengue and Zika virus recombinant envelope proteins
- Author
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Ashutosh Tripathy, James A. Brackbill, Michael J. Miley, Stephan T. Kudlacek, Alexander Matthew Payne, Lakshmanane Premkumar, Stephen D Graham, Andrey A. Bobkov, Stefan W. Metz, Aravinda M. de Silva, and Brian Kuhlman
- Subjects
0301 basic medicine ,Protein subunit ,Dimer ,Biochemistry ,Microbiology ,Epitope ,Dengue fever ,Zika virus ,law.invention ,Body Temperature ,Dengue ,03 medical and health sciences ,chemistry.chemical_compound ,Viral Envelope Proteins ,law ,medicine ,Humans ,Molecular Biology ,chemistry.chemical_classification ,030102 biochemistry & molecular biology ,biology ,Chemistry ,Protein Stability ,Zika Virus Infection ,Viral Vaccines ,Cell Biology ,Zika Virus ,Dengue Virus ,medicine.disease ,biology.organism_classification ,Virology ,Recombinant Proteins ,030104 developmental biology ,Vaccines, Subunit ,Recombinant DNA ,biology.protein ,Antibody ,Protein Multimerization ,Glycoprotein - Abstract
The spread of dengue (DENV) and Zika virus (ZIKV) is a major public health concern. The primary target of antibodies that neutralize DENV and ZIKV is the envelope (E) glycoprotein, and there is interest in using soluble recombinant E (sRecE) proteins as subunit vaccines. However, the most potent neutralizing antibodies against DENV and ZIKV recognize epitopes on the virion surface that span two or more E proteins. Therefore, to create effective DENV and ZIKV vaccines, presentation of these quaternary epitopes may be necessary. The sRecE proteins from DENV and ZIKV crystallize as native-like dimers, but studies in solution suggest that these dimers are marginally stable. To better understand the challenges associated with creating stable sRecE dimers, we characterized the thermostability of sRecE proteins from ZIKV and three DENV serotypes, DENV2-4. All four proteins irreversibly unfolded at moderate temperatures (46-53 °C). At 23 °C and low micromolar concentrations, DENV2 and ZIKV were primarily dimeric, and DENV3-4 were primarily monomeric, whereas at 37 °C, all four proteins were predominantly monomeric. We further show that the dissociation constant for DENV2 dimerization is very temperature-sensitive, ranging from
- Published
- 2018