Use of Circular Dichroism in the characterization of the fusion protein SARS-CoV-2 S protein (RBD)-hFc
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Ingrid Ruíz
Abstract
From the receptor-binding domain (RBD) of the SARS-CoV-2 virus, which causes coronavirus disease 2019 (COVID-19), a RBD-hFc fusion protein was obtained at the Center of Molecular Immunology (Havana, Cuba). This fusion protein was used in the construction of a diagnostic device for COVID-19 called Ultramicroenzyme-Linked Immunosorbent Assay (UMELISA)-SARS-CoV-2-IgG and it is currently been used in the studies of biological activity of the Cuban vaccine Abdala (CIGB-66). In this work, Circular Dichroism (CD) is used to characterize this protein. Using Far Ultraviolet Circular Dichroism (FAR-UV CD), it was determined that the protein has a secondary structure in the form of a sheet-β fundamentally. Using this technique, a thermodynamic study was carried out and it was determined that the melting temperature (Tm) of the protein is 71.5 °C. Information about the tertiary structure of the protein was obtained using Near Ultraviolet Circular Dichroism (NEAR-UV CD) and Molecular Fluorescence; they indicates that the protein has a three-dimensional folding associated with the aromatic amino acids in its structure, where tryptophan (Trp) is located inside the folded structure of the protein while tyrosine (Tyr) is exposed to the solvent.
Article note:
A collection of invited papers based on presentations at the Virtual Conference on Chemistry and its Applications (VCCA-2021) held on-line, 9–13 August 2021.
Secondary structure and its content in % of the RBD-hFc protein spectra obtained at a temperature range of 20–95 °C, with intervals of 5 °C (Results of deconvolution analysis of the spectra in Figure 5 using the BeStSel).
| Temperature | α-helix | β-sheet | Random coil |
|---|---|---|---|
| 20 | 5.1 | 49.9 | 45.0 |
| 25 | 7.4 | 50.1 | 42.5 |
| 30 | 5.8 | 49.9 | 44.3 |
| 35 | 5.7 | 49.1 | 45.2 |
| 40 | 8.3 | 48.6 | 43.1 |
| 45 | 6.4 | 51 | 42.6 |
| 50 | 7.5 | 52.3 | 40.2 |
| 55 | 4.2 | 51 | 44.8 |
| 60 | 5.6 | 51.5 | 42.9 |
| 65 | 8.9 | 47.6 | 43.5 |
| 70 | 10.4 | 43.7 | 45.9 |
| 75 | 5.1 | 45.4 | 49.5 |
| 80 | 5.9 | 45.7 | 48.4 |
| 85 | 11.3 | 44.2 | 44.5 |
| 90 | 7.1 | 46.5 | 46.4 |
| 95 | 6.8 | 45.6 | 47.6 |
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Artikel in diesem Heft
- Frontmatter
- In this issue
- Editorial
- The virtual conference on chemistry and its applications, VCCA-2021, 9–13 August 2021
- Conference papers
- Hexabenzocoronene functionalized with antiaromatic S- and Se-core-modified porphyrins (isophlorins): comparison with the dyad with regular porphyrin
- Bonding analysis of the C2 precursor Me3E–C2–I(Ph)FBF3 (E = C, Si, Ge)
- Supporting the fight against the proliferation of chemical weapons through cheminformatics
- Disinfecting activity of some diphenyltin(IV) benzoate derivative compounds
- HCV genotype-specific drug discovery through structure-based virtual screening
- ExcelAutomat 1.4: generation of supporting information
- Use of Circular Dichroism in the characterization of the fusion protein SARS-CoV-2 S protein (RBD)-hFc
- Experimental determination of activation rate constant and equilibrium constant for bromo substituted succinimide initiators for an atom transfer radical polymerization process
- Degradation of o-, m-, p-cresol isomers using ozone in the presence of V2O5-supported Mn, Fe, and Ni catalysts
- The beginnings of chemistry: from ancient times until 1661
- Chemical substitution in processes for inherently safer design: pros and cons
- Experimental and theoretical study of the dye-sensitized solar cells using Hibiscus sabdariffa plant pigment coupled with polyaniline/graphite counter electrode
Artikel in diesem Heft
- Frontmatter
- In this issue
- Editorial
- The virtual conference on chemistry and its applications, VCCA-2021, 9–13 August 2021
- Conference papers
- Hexabenzocoronene functionalized with antiaromatic S- and Se-core-modified porphyrins (isophlorins): comparison with the dyad with regular porphyrin
- Bonding analysis of the C2 precursor Me3E–C2–I(Ph)FBF3 (E = C, Si, Ge)
- Supporting the fight against the proliferation of chemical weapons through cheminformatics
- Disinfecting activity of some diphenyltin(IV) benzoate derivative compounds
- HCV genotype-specific drug discovery through structure-based virtual screening
- ExcelAutomat 1.4: generation of supporting information
- Use of Circular Dichroism in the characterization of the fusion protein SARS-CoV-2 S protein (RBD)-hFc
- Experimental determination of activation rate constant and equilibrium constant for bromo substituted succinimide initiators for an atom transfer radical polymerization process
- Degradation of o-, m-, p-cresol isomers using ozone in the presence of V2O5-supported Mn, Fe, and Ni catalysts
- The beginnings of chemistry: from ancient times until 1661
- Chemical substitution in processes for inherently safer design: pros and cons
- Experimental and theoretical study of the dye-sensitized solar cells using Hibiscus sabdariffa plant pigment coupled with polyaniline/graphite counter electrode
