SARS-CoV-2 Spike Protein Set (LMNG, DIBMA Glycerol & Nanodisc Complex)

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Description

SPIKE stabilized in a SMALP copolymer nanodisc and depiction of its individual domains — **Figure 1:** Overview of SPIKE's domains and overall structure. Note that the protein is a homo-trimer. Using either LMNG detergent, MSP nanodiscs or synthetic copolymer nanodiscs, that trimer remains intact.

The coronavirus, also known as SARS-CoV-2, enters the cell by using its surface SPIKE. SPIKE is processed on the cell's surface by TMPRSS2, a serine protease. It then subsequently binds to ACE2 a cell surface receptor.

The Native SPIKE protein is a trimer that is located in the coronavirus membrane. Therefore to get pure & native SPIKE this trimer needs to be kept intact. And Cube Biotech offers this intact form of SPIKE. Figure 1 highlights how this stabilization is achieved and provides insights into the overall structure of SPIKE.

THis 3 in one protein set contains the following SPIKE protein products of Cube Biotech:

Datasheets

Feature - SPIKE in LMNG
Alternative names	SPIKE_SARS2 Spike glycoprotein
UniProt Number	P0DTC2
Protein class	Single span transmembrane protein
Organism	Severe acute respiratory syndrome coronavirus 2 (2019-nCoV) (SARS-CoV-2)
Sequence, explained	Full-length sequence (aa 1 – 1273), furin cleavage site “RRAR” mutated to “GSAG”; C-terminal Rho1D4 tag fused with spacer “GSSG” to protein sequence
Affinity tags	C-terminal Rho1D4-tag
Expression Host	Hek293 Expi cells
Size	1286 amino acids (including Rho1D4-tag and linker) 142114 Da
Buffer composition	20 mM Hepes pH 7.5; 150 mM NaCl, 0.001 % LMNG
Purity	>98%, determined via SDS-PAGE (Figure 2 A and B)
Homogeneity	Native Gel, see Fig. 2 C
Purified via	PureCube Rho1D4 Agarose
Function	Host cell surface receptor binding; fusion of virus membrane with host endosome membrane
Shipping conditions	Dry Ice
Storage conditions	–80°C. Avoid freeze-thaw cycles

Feature - SPIKE in MSP nanodisc
Alternative names	SPIKE_SARS2 Spike glycoprotein
UniProt Number	P0DTC2
Protein class	Single span transmembrane protein
Organism	Severe acute respiratory syndrome coronavirus 2 (2019-nCoV) (SARS-CoV-2)
Sequence, explained	Full-length sequence (aa 1 – 1273), furin cleavage site “RRAR” mutated to “GSAG”; C-terminal Rho1D4 tag fused with spacer “GSSG” to protein sequence
Affinity tags	C-terminal Rho1D4-tag
Expression Host	Hek293 Expi cells
Size	1286 amino acids (including Rho1D4-tag and linker) 142114 Da
Buffer composition	20 mM Hepes pH 7.5; 150 mM NaCl,
Stabilizing MSP nanodisc	MSP1D1-His, assembly with POPC lipids (Membrane scaffold protein based nanodisc; MSP1D1 = 25,3 kDa)
Purity	>98%, determined via SDS-PAGE (Figure 3 and 4)
Homogeneity	Native Gel, see Fig. 3 and 4
Purified via	PureCube Rho1D4 Agarose
Function	Host cell surface receptor binding; fusion of virus membrane with host endosome membrane
Shipping conditions	Dry Ice
Storage conditions	–80°C. Avoid freeze-thaw cycles

Feature - SPIKE in DIBMA Glycerol
Alternative names	SPIKE_SARS2 Spike glycoprotein
UniProt Number	P0DTC2
Protein class	Single span transmembrane protein
Organism	Severe acute respiratory syndrome coronavirus 2 (2019-nCoV) (SARS-CoV-2)
Sequence, explained	Full-length sequence (aa 1 – 1273), furin cleavage site “RRAR” mutated to “GSAG”; C-terminal Rho1D4 tag fused with spacer “GSSG” to protein sequence
Affinity tags	C-terminal Rho1D4-tag
Expression Host	Hek293 Expi cells
Size	1286 amino acids (including Rho1D4-tag and linker) 142114 Da
Buffer composition	20 mM Hepes pH 7.5; 150 mM NaCl, buffer does not contain free DIBMA Glycerol (DIBMA: Diisobutylene-maleic acid)
Stabilization agent	DIBMA Glycerol
Purity	>98%, determined via SDS-PAGE (Figure 5 A and B)
Homogeneity	Native Gel, see Fig. 5 C
Purified via	PureCube Rho1D4 Agarose
Function	Host cell surface receptor binding; fusion of virus membrane with host endosome membrane
Shipping conditions	Dry Ice
Storage conditions	–80°C. Avoid freeze-thaw cycles

Lab Results

SPIKE (wt) SDS-PAGE — **Figure 2:** Data from our lab regarding SPIKE stabilized in a detergent mycelle. A: SDS-Page of purified SPIKE in detergent mycelle. B: Western blot of Rho1d4-tagged SPIKE, originated from the SDS Page seen in "A". C: Native protein Gel of purified and stabilized SPIKE in detergent mycelles.

SPIKE (wt) MSP nanodisc SDS-PAGE — **Figure 3:** SDS Page of purified SPIKE protein in MSP1D1 nanodiscs. The band that represents the MSP protein can also be seen.

MSP nanodisc SPIKE gel filtration — **Figure 4:** Gel filtration result of SPIKE in MSP nanodisc. The peak that represents spike is marked with a red arrow.

SDS-PAge of SPIKE protein in Copolymer DIMBA Glycerol nanodisc — **Figure 5:** Data from our lab regarding SPIKE stabilized and purified in synthetic nanodiscs (Dibma Glycerol). A: SDS-PAGE of SPIKE in synthetic nanodisc. B: Western blot of Rhot1d4 tagged SPIKE, originated from the SDS Page seen in "A" C: Native protein Gel of purified and stabilized SPIKE in synthetic copolymer nanodisc.

FAQ

Can I get the datasheets for the SPIKE (wt) proteins of this set?

Yes, just download them here.

Can I infect myself with COVID-19 while working with this product? With the product being the source of the infection.

No, this is the isolated surface protein of the virus. Not a complete and harmful version of SARS-CoV-2.

Do you also offer other SPIKE variants / mutations?

Yes, we do! Look at this LIST.

Do you also offer other full-length membrane proteins?

Yes, we do. Just browse through our product protofolio. In case we do not have your desired membrane protein in our store, send a request to our protein services.