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TitleSARS-CoV-2 S2P spike ages through distinct states with altered immunogenicity.
Journal, issue, pagesJ Biol Chem, Vol. 297, Issue 4, Page 101127, Year 2021
Publish dateAug 27, 2021
AuthorsAdam S Olia / Yaroslav Tsybovsky / Steven J Chen / Cuiping Liu / Alexandra F Nazzari / Li Ou / Lingshu Wang / Wing-Pui Kong / Kwan Leung / Tracy Liu / Tyler Stephens / I-Ting Teng / Shuishu Wang / Eun Sung Yang / Baoshan Zhang / Yi Zhang / Tongqing Zhou / John R Mascola / Peter D Kwong /
PubMed AbstractThe SARS-CoV-2 spike is the primary target of virus-neutralizing antibodies and critical to the development of effective vaccines against COVID-19. Here, we demonstrate that the prefusion-stabilized ...The SARS-CoV-2 spike is the primary target of virus-neutralizing antibodies and critical to the development of effective vaccines against COVID-19. Here, we demonstrate that the prefusion-stabilized two-proline "S2P" spike-widely employed for laboratory work and clinical studies-unfolds when stored at 4 °C, physiological pH, as observed by electron microscopy (EM) and differential scanning calorimetry, but that its trimeric, native-like conformation can be reacquired by low pH treatment. When stored for approximately 1 week, this unfolding does not significantly alter antigenic characteristics; however, longer storage diminishes antibody binding, and month-old spike elicits virtually no neutralization in mice despite inducing high ELISA-binding titers. Cryo-EM structures reveal the folded fraction of spike to decrease with aging; however, its structure remains largely similar, although with varying mobility of the receptor-binding domain. Thus, the SARS-CoV-2 spike is susceptible to unfolding, which affects immunogenicity, highlighting the need to monitor its integrity.
External linksJ Biol Chem / PubMed:34461095 / PubMed Central
MethodsEM (single particle)
Resolution2.6 - 3.5 Å
Structure data

23982

7mtc

EMDB-23982, PDB-7mtc:
Structure of freshly purified SARS-CoV-2 S2P spike at pH 7.4
Method: EM (single particle) / Resolution: 2.6 Å

23983

7mtd

EMDB-23983, PDB-7mtd:
Structure of aged SARS-CoV-2 S2P spike at pH 7.4
Method: EM (single particle) / Resolution: 3.5 Å

23984

7mte

EMDB-23984, PDB-7mte:
Structure of SARS-CoV-2 S2P spike at pH 7.4 refolded by low-pH treatment
Method: EM (single particle) / Resolution: 3.2 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

Source
  • severe acute respiratory syndrome coronavirus 2
KeywordsVIRAL PROTEIN / COVID-19 / SARS-CoV-2 spike / S2P

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