@article{fdi:010053097,
  title = {{C}omparative production analysis of three {P}hlebovirus nucleoproteins under denaturing or non-denaturing conditions for crystallographic studies},
  author = {{L}antez, {V}. and {D}alle, {K}. and {C}harrel, {R}. and {B}aronti, {C}{\'e}cile and {C}anard, {B}. and {C}outard, {B}.},
  editor = {},
  language = {{ENG}},
  abstract = {{N}ucleoproteins ({NP}s) encapsidate the {P}hlebovirus genomic (-){RNA}. {U}pon recombinant expression, {NP}s tend to form heterogeneous oligomers impeding characterization of the encapsidation process through crystallographic studies. {T}o overcome this problem, we set up a standard protocol in which production under both non-denaturing and denaturing/ refolding conditions can be investigated and compared. {T}he protocol was applied for three phlebovirus {NP}s, allowing an optimized production strategy for each of them. {R}emarkably, the {R}ift {V}alley fever virus {NP} was purified as a trimer under native conditions and yielded protein crystals whereas the refolded version could be purified as a dimer. {Y}ields of trimeric {T}oscana virus {NP} were higher from denaturing than from native condition and lead to crystals. {T}he production of {S}andfly {F}ever {S}icilian virus {NP} failed in both protocols. {T}he comparative protocols described here should help in rationally choosing between denaturing or non-denaturing conditions, which would finally result in the most appropriate and relevant oligomerized protein species. {T}he structure of the {R}ift {V}alley fever virus {NP} has been recently published using a refolded monomeric protein and we believe that the process we devised will contribute to shed light in the genome encapsidation process, a key stage in the viral life cycle.},
  keywords = {},
  booktitle = {},
  journal = {{P}los {N}eglected {T}ropical {D}iseases},
  volume = {5},
  numero = {1},
  pages = {e936},
  ISSN = {1935-2727},
  year = {2011},
  DOI = {10.1371/journal.pntd.0000936},
  URL = {https://www.documentation.ird.fr/hor/fdi:010053097},
}