RRC ID 66154
著者 Ohki S, Imamura T, Higashimura Y, Matsumoto K, Mori M.
タイトル Similarities and differences in the conformational stability and reversibility of ORF8, an accessory protein of SARS-CoV-2, and its L84S variant.
ジャーナル Biochem Biophys Res Commun
Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), has the characteristic accessory protein ORF8. Although clinical reports indicate that ORF8 variant strains (Δ382 and L84S variants) are less likely to cause severe illness, functional differences between wild-type and variant ORF8 are unknown. Furthermore, the physicochemical properties of the ORF8 protein have not been analyzed. In this study, the physicochemical properties of the wild-type ORF8 and its L84S variant were analyzed and compared. Using the tobacco BY-2 cell production system, which has been successfully used to produce the wild-type ORF8 protein with a single conformation, was used to successfully produce the ORF8 L84S variant protein at the same level as wild-type ORF8. The produced proteins were purified, and their temperature and pH dependencies were examined using nuclear magnetic resonance spectra. Our data suggested that the wild-type and L84S variant ORF8 structures are highly stable over a wide temperature range. Both proteins displayed an aggregated conformation at higher temperature that reverted when the temperature was decreased to room temperature. Moreover, ORF8 precipitated at acidic pH and this precipitation was reversed when the solution pH was shifted to neutral. Interestingly, the L84S variant exhibited greater solubility than wild-type ORF8 under acidic conditions. Thus, the finding indicated that conformational stability and reversibility of ORF8 are key properties related to function in oppressive environments.
巻・号 563
ページ 92-97
公開日 2021-7-23
DOI 10.1016/j.bbrc.2021.05.074
PII S0006-291X(21)00857-3
PMID 34062392
PMC PMC8149210
MeSH COVID-19 / metabolism COVID-19 / pathology COVID-19 / virology* Humans Molecular Conformation Mutation Nuclear Magnetic Resonance, Biomolecular / methods Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / isolation & purification SARS-CoV-2 / chemistry* SARS-CoV-2 / isolation & purification SARS-CoV-2 / metabolism Structure-Activity Relationship Viral Proteins / chemistry* Viral Proteins / genetics Viral Proteins / metabolism
IF 2.985
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 rpc00001