Recombinant SARS-CoV-2 C.1.2 S (GCN4-IZ) His-tag Protein, CF

Recombinant SARS-CoV-2 C.1.2 Spike (GCN4-IZ) His-tag Protein (Catalog # 11007-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

2 μg/lane of Recombinant SARS-CoV-2 C.1.2 Spike (GCN4-IZ) His-tag Protein (Catalog # 11007-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 142-170 kDa.

• Reactivity: V
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant SARS-CoV-2 C.1.2 S (GCN4-IZ) His-tag Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag (Catalog # (Catalog # 933-ZN).
• Source: Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein
  

  SARS-CoV-2 C.1.2 S (Val16-Lys1211) (Pro25Leu, Cys136Phe, Tyr144del, Arg190Ser, Asp215Gly, Ala243del, Leu244del, Tyr449His, Glu484Lys, Asn501Tyr, Leu585Phe, Asp614Gly, His655Tyr, Asn679Lys, Thr716Ile, Thr859Asn) (Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro) Accession # YP_009724390.1	GCN4-IZ	6-His tag
  N-terminus		C-terminus

• Accession #: YP_009724390.1
• N-terminal Sequence: Val16
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 138 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 142-170 kDa, under reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant SARS-CoV-2 C.1.2 S (GCN4-IZ) His-tag Protein, CF

• 2019-nCoV S Protein
• 2019-nCoV Spike
• COVID-19 Spike
• E2
• Human coronavirus spike glycoprotein
• Peplomer protein
• S glycoprotein
• S Protein
• SARS-COV-2 S protein
• SARS-COV-2 Spike glycoprotein
• SARSCOV2 Spike protein
• SARS-CoV-2
• Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein
• Spike glycoprotein
• Spike
• surface glycoprotein

Background

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that also include MERS‑CoV and SARS-CoV-1. Coronaviruses are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M) and Nucleocapsid protein (N) (1). The SARS-CoV-2 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 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 sequence identity with S protein of SARS‑CoV-1 and MERS, respectively (6, 7). The low aa sequence homology is consistent with the finding that SARS and MERS‑CoV bind different cellular receptors (8). The RBD of SARS-CoV-2 binds a metallopeptidase, angiotensin-converting enzyme 2 (ACE‑2), similar to SARS-CoV-1, but with much higher affinity and faster binding kinetics (9). Before binding to the ACE‑2 receptor, structural analysis of the S1 trimer shows that only one of the three RBD domains is in the "up" conformation. This is an unstable and transient state that passes between trimeric subunits but is nevertheless an exposed state to be targeted for neutralizing antibody therapy (10). Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE‑2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (11). There is also promising work showing that the RBD may be used to detect presence of neutralizing antibodies present in a patient's bloodstream, consistent with developed immunity after exposure to the SARS-CoV-2 (12). A SARS-CoV-2 variant (C.1.2) carrying the aa substitution Pro25Leu, Cys136Phe, Tyr144del, Arg190Ser, Asp215Gly, Ala243del, Leu244del, Tyr449His, Glu484Lys, Asn501Tyr, Leu585Phe, Asp614Gly, His655Tyr, Asn679Lys, Thr716Ile, and Thr859Asn in spike protein was evolved from C.1. It was first identified in May 2021 in South Africa and rapidly spreaded globally. Whether these mutations in the spike protein would cause more severe symptom or decrease the efficacy of vaccine-induced immunity is still under investigation (13).

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. Li, W. et al. (2003) Nature 426:450.
7. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
8. Jiang, S. et al. (2020) Trends. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
9. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
10. Wrapp, D. et al. (2020) Science 367:1260.
11. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.1.
12. Okba, N.M.A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
13. Scheepers, C. et al. (2021) medRxiv https://doi.org/10.1101/2021.08.20.21262342.

Bioinformatics

• Uniprot:
  • YP_009724390.1(Llama)