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Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein, CF

Catalog # 10561-CV | R&D Systems, Inc. a Bio-Techne Brand
GCN4-IZ trimerization domain, Stabilized Prefusion Conformation, Resistant to Furin Cleavage
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10561-CV-100

Key Product Details

Source

HEK293

Accession #

Structure / Form

Non-covalent trimer / multimer

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein
SARS-CoV-2 Spike
(Val16-Lys1211)
(R682S)(R685S)(K986P)(V987P)
Accession # YP_009724390.1
GCN4-IZ HHHHHH
N-terminus C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Val16

Predicted Molecular Mass

138 kDa

SDS-PAGE

144-175 kDa, under reducing conditions

Activity

Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag  (Catalog # 933-ZN).

Scientific Data Images for Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein, CF

Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein SDS-PAGE

Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein SDS-PAGE

2 μg/lane of Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein (Catalog # 10561-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 144-175 kDa.
Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein Binding Activity

Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein Binding Activity

Recombinant SARS-CoV-2 Spike (GCN4-IZ) His-tag Protein (Catalog # 10561-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.
Surface plasmon resonance (SPR) sensorgram of Human ACE-2 binding to SARS-CoV-2 Spike protein (GCN4-IZ)

Binding of ACE-2 to SARS-CoV-2 Spike protein by surface plasmon resonance (SPR).

Recombinant SARS-CoV-2 Spike protein with a GCN4-IZ domain and His-tag was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.37 nM and 94.3 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=2.580 nM. (Biacore T200).

Formulation, Preparation and Storage

10561-CV
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Spike

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). SARS-CoV-2 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. The S protein is homotrimeric, with each ~180-kDa monomer consisting of two subunits, S1 and S2 (2). In SARS-CoV-2, as with most coronaviruses, proteolytic cleavage of the S protein into the S1 and S2 subunits is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (3-5). The S protein of SARS-CoV-2 shares 75% and 29% amino acid (aa) sequence identity with the S protein of SARS-CoV-1 and MERS, respectively. The S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds Angiotensin-Converting Enzyme 2 (ACE2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1 (6). Based on structural biology studies, the RBD can be oriented either in the up/standing or down/lying state with the up/standing state associated with higher pathogenicity (7). Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (8). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (9, 10). A SARS-CoV-2 variant carrying the S protein aa change D614G has become the most prevalent form in the global pandemic and has been associated with greater infectivity and higher viral load (11, 12).

References

  1. Wu, F. et al. (2020) Nature 579:265.
  2. Tortorici, M.A. and D. Veesler (2019). Adv. Virus Res. 105:93.
  3. Bosch, B.J. et al. (2003). J. Virol. 77:8801.
  4. Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
  5. Millet, J.K. and G.R. Whittaker (2015) Virus Res. 202:120.
  6. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  7. Yuan, Y. et al. (2017) Nat. Commun. 8:15092.
  8. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
  9. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
  10. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.
  11. Korber, B. et al. (2020) Cell 182, 812.
  12. Zhang, L. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.06.12.148726v1.

Long Name

Spike Protein

Alternate Names

S Protein

Entrez Gene IDs

918758 (HCoV-229E); 2943499 (HCoV-NL63); 39105218 (HCoV-OC43); 37616432 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

Product Documents for Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant SARS-CoV-2 Spike (GCN4-IZ) His Protein, CF

For research use only

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