Thermodynamic stability of domain III from the envelope protein of flaviviruses and its improvement by molecular design

Nora Zidane, Philippe Dussart, Laetitia Bremand, Maria Elena Villani, Hugues Bedouelle

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The Flavivirus genus includes widespread and severe human pathogens like the four serotypes of dengue virus (DENV1 to DENV4), yellow fever virus, Japanese encephalitis virus and West Nile virus. Domain III (ED3) of the viral envelope protein interacts with cell receptors and contains epitopes recognized by virus neutralizing antibodies. Its structural, antigenic and immunogenic properties have been thoroughly studied contrary to its physico-chemical properties. Here, the ED3 domains of the above pathogenic flaviviruses were produced in the periplasm of Escherichia coli. Their thermodynamic stabilities were measured and compared in experiments of unfolding equilibriums, induced with chemicals or heat and monitored through protein fluorescence. A designed ED3 domain, with the consensus sequence of DENV strains from all serotypes, was highly stable. The low stability of the ED3 domain from DENV3 was increased by three changes of residues in the protein core without affecting its reactivity towards DENV-infected human serums. Additional changes showed that the stability of ED3 varied with the DENV3 genotype. The Tmof ED3 was higher than 69°C for all the tested viruses and reached 86°C for the consensus ED3. The latter, deprived of its disulfide bond by mutations, was predominantly unfolded at 20°C. These results will help better understand and design the properties of ED3 for its use as diagnostic, vaccine or therapeutic tools. © 2013 The Author.
Original languageEnglish
Pages (from-to)389 - 399
Number of pages11
JournalProtein Engineering, Design and Selection
Issue number6
Publication statusPublished - Jun 2013


All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Molecular Biology

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