Name: Recombinant Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein (S), partial,Biotinylated (Active)
Brand: CUSABIO
SKU: CSB-MP3324GMY1-B

Product Details

Purity

Greater than 90% as determined by SDS-PAGE.
Greater than 95% as determined by SEC-HPLC.

Endotoxin

Less than 1.0 EU/ug as determined by LAL method.

Activity

Measured by its binding ability in a functional ELISA. Immobilized human ACE2 at 2 μg/ml can bind Biotinylated-SARS-CoV-2-S1-RBD (CSB-MP3324GMY1-B), the EC₅₀ is 5.087-7.050 ng/ml.

Target Names

Uniprot No.

P0DTC2

Research Area

Others

Alternative Names

S; 2; Spike glycoprotein; S glycoprotein; E2; Peplomer protein)

Species

Severe acute respiratory syndrome coronavirus 2 (2019-nCoV) (SARS-CoV-2)

Source

Mammalian cell

Expression Region

319-541aa

Target Protein Sequence

RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF

Mol. Weight

54.1 kDa

Protein Length

Partial

Tag Info

C-terminal mFC-Avi-tagged

Form

Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

Lyophilized from a 0.2 μm sterile filtered PBS, 6% Trehalose, pH 7.4.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

Recombinant severe acute respiratory syndrome coronavirus 2 Spike glycoprotein (S) production involves several steps starting with the isolation of the target gene that corresponds to the 319-541aa of the SARS-CoV-2 S protein. The gene is fused with a C-terminal mFC-Avi-tag gene and then cloned into an expression vector and introduced into mammalian cells via transformation. The mammalian cells express the recombinant protein, which is harvested from the cell lysate. The protein is purified using affinity chromatography. The final step involves validating the protein's purity and functionality. Its purity is determined through SDS-PAGE and SEC-HPLC, reaching over 90% and 95%, respectively. Its endotoxin content is less than 1.0 EU/ug as determined by the LAL method. This biotinylated recombinant SARS-CoV-2 S protein has been validated as an active protein in a functional ELISA.

The S protein of SARS-CoV-2 is a crucial component of the virus responsible for mediating host-cell entry. It is a type-1 transmembrane glycoprotein composed of two subunits, S1 and S2, responsible for attachment and entry of the virus, respectively [1][2]. The S protein is highly glycosylated and projects from the viral surface, facilitating attachment to the ACE2 receptor on host cells [3]. This protein is the main target for vaccine development and clinical diagnosis due to its antigenicity and role in virus entry [4][5]. Additionally, the S protein plays a key role in syncytia formation and virus entry by associating with the ACE2 receptor [5].

References:
[1] Y. Huang, C. Yang, X. Xu, W. Xü, & S. Liu, Structural and functional properties of sars-cov-2 spike protein: potential antivirus drug development for covid-19, Acta Pharmacologica Sinica, vol. 41, no. 9, p. 1141-1149, 2020. https://doi.org/10.1038/s41401-020-0485-4
[2] M. Hussain, N. Jabeen, A. Amanullah, B. Aa, B. Aziz, S. Shabbiret al., Structural basis of sars-cov-2 spike protein priming by tmprss2,, 2020. https://doi.org/10.1101/2020.04.21.052639
[3] U. Shekhawat and A. Chowdhury, Computational and comparative investigation of hydrophobic profile of spike protein of sars-cov-2 and sars-cov, Journal of Biological Physics, vol. 48, no. 4, p. 399-414, 2022. https://doi.org/10.1007/s10867-022-09615-x
[4] J. Zhou, J. Sun, G. Lu, W. Wang, & W. Lin, Sars-cov-2 spike protein evolution may cause difficulties for vaccine,, 2020. https://doi.org/10.21203/rs.3.rs-80010/v1
[5] D. Li, Y. Liu, Y. Lu, S. Gao, & L. Zhang, Palmitoylation of sars‐cov‐2 s protein is critical for s‐mediated syncytia formation and virus entry, Journal of Medical Virology, vol. 94, no. 1, p. 342-348, 2021. https://doi.org/10.1002/jmv.27339

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Target Background

Function

attaches the virion to the cell membrane by interacting with host receptor, initiating the infection. Binding to human ACE2 receptor and internalization of the virus into the endosomes of the host cell induces conformational changes in the Spike glycoprotein. Binding to host NRP1 and NRP2 via C-terminal polybasic sequence enhances virion entry into host cell. This interaction may explain virus tropism of human olfactory epithelium cells, which express high level of NRP1 and NRP2 but low level of ACE2. The stalk domain of S contains three hinges, giving the head unexpected orientational freedom. Uses human TMPRSS2 for priming in human lung cells which is an essential step for viral entry. Can be alternatively processed by host furin. Proteolysis by cathepsin CTSL may unmask the fusion peptide of S2 and activate membranes fusion within endosomes.; mediates fusion of the virion and cellular membranes by acting as a class I viral fusion protein. Under the current model, the protein has at least three conformational states: pre-fusion native state, pre-hairpin intermediate state, and post-fusion hairpin state. During viral and target cell membrane fusion, the coiled coil regions (heptad repeats) assume a trimer-of-hairpins structure, positioning the fusion peptide in close proximity to the C-terminal region of the ectodomain. The formation of this structure appears to drive apposition and subsequent fusion of viral and target cell membranes.; Acts as a viral fusion peptide which is unmasked following S2 cleavage occurring upon virus endocytosis.; May down-regulate host tetherin (BST2) by lysosomal degradation, thereby counteracting its antiviral activity.

Gene References into Functions

Study presents crystal structure of C-terminal domain of SARS-CoV-2 (SARS-CoV-2-CTD) spike S protein in complex with human ACE2 (hACE2); hACE2-binding mode similar overall to that observed for SARS-CoV. However, details at the binding interface show that key residue substitutions in SARS-CoV-2-CTD slightly strengthen the interaction and lead to higher affinity for receptor binding than SARS-CoV receptor-binding domain. PMID: 32378705
crystal structure of the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 bound to the cell receptor ACE2 PMID: 32365751
crystal structure of the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (engineered to facilitate crystallization) in complex with ACE2 PMID: 32320687
Out of the two isolates from India compared to the isolates from Wuhan, China, one was found to harbor a mutation in its receptor-binding domain (RBD) at position 407 where, arginine was replaced by isoleucine. This mutation has been seen to change the secondary structure of the protein at that region and this can potentially alter receptor binding of the virus. PMID: 32275855
Structural modeling of the SARS-CoV-2 spike glycoprotein show similar receptor utilization between SARS-CoV-2 and SARS-CoV, despite a relatively low amino acid similarity in the receptor binding module. Compared to SARS-CoV and all other coronaviruses in Betacoronavirus lineage B, an extended structural loop containing basic amino acids were identified at the interface of the receptor binding (S1) and fusion (S2) domains. PMID: 32245784
crystal structure of CR3022, a neutralizing antibody from a SARS patient, in complex with the receptor-binding domain of the SARS-CoV-2 spike (S) protein to 3.1 A; study provides insight into how SARS-CoV-2 can be targeted by the humoral immune response and revealed a conserved, but cryptic epitope shared between SARS-CoV-2 and SARS-CoV PMID: 32225176
SARS-CoV and SARS-CoV-2 spike proteins have comparable binding affinities achieved by balancing energetics and dynamics. The SARS-CoV-2-ACE2 complex contains a higher number of contacts, a larger interface area, and decreased interface residue fluctuations relative to the SARS-CoV-ACE2 complex. PMID: 32225175
Interaction interface between cat/dog/pangolin/Chinese hamster ACE2 and SARS-CoV/SARS-CoV-2 S protein was simulated through homology modeling. Authors identified that N82 of ACE2 showed closer contact with receptor-binding domain of S protein than human ACE2. PMID: 32221306
SARS-CoV-2 S glycoprotein harbors a furin cleavage site at the boundary between the S1/S2 subunits, which is processed during biogenesis and sets this virus apart from SARS-CoV and SARS-related CoVs; determined cryo-EM structures of the SARS-CoV-2 S ectodomain trimer. PMID: 32201080
Study demonstrates that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. PMID: 32155444
The ACE2-B0AT1 complex exists as a dimer of heterodimers. Structural alignment of the RBD-ACE2-B0AT1 ternary complex with the S protein of SARS-CoV-2 suggests that two S protein trimers can simultaneously bind to an ACE2 homodimer. PMID: 32142651
study demonstrated SARS-CoV-2 S protein entry on 293/hACE2 cells is mainly mediated through endocytosis, and PIKfyve, TPC2 and cathepsin L are critical for virus entry; found that SARS-CoV-2 S protein could trigger syncytia in 293/hACE2 cells independent of exogenous protease; there was limited cross-neutralization activity between convalescent sera from SARS and COVID-19 patients PMID: 32132184
study determined a 3.5-angstrom-resolution cryo-electron microscopy structure of the 2019-nCoV S trimer in the prefusion conformation; provided biophysical and structural evidence that the 2019-nCoV S protein binds angiotensin-converting enzyme 2 (ACE2) with higher affinity than does severe acute respiratory syndrome (SARS)-CoV S PMID: 32075877

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Subcellular Location

Virion membrane; Single-pass type I membrane protein. Host endoplasmic reticulum-Golgi intermediate compartment membrane; Single-pass type I membrane protein. Host cell membrane; Single-pass type I membrane protein.

Protein Families

Betacoronaviruses spike protein family

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Code	CSB-MP3324GMY1-B
MSDS	MSDS
Size	$456
	Order now
Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel. Activity Measured by its binding ability in a functional ELISA. Immobilized human ACE2 at 2 μg/ml can bind Biotinylated-SARS-CoV-2-S1-RBD (CSB-MP3324GMY1-B), the EC₅₀ is 5.087-7.050 ng/ml The purity of S was greater than 95% as determined by SEC-HPLC
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Recombinant Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein (S), partial,Biotinylated (Active)

Product Details

Citations

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