Product Information
Purity: > 85% as determined by SDS-PAGE
Endotoxin: <1.0 EU per μg protein as determined by the LAL method.
Activity: Tests in progress
Protein construction
A DNA sequence encoding Zaire Ebola virus (H. sapiens-wt / GIN / 2014 / Kissidougou-C15 strain) GP (AHX24649.1) (Met1-Gln650) was expressed with a polyhistidine tag at the C-terminal end.
Registration number: AHX24649.1
Expressed Host: Baculovirus- Insect Cells
Species: EBOV
Molecular mass
Recombinant Zaire Ebola virus (H. sapiens-wt / GIN / 2014 / Kissidougou-C15 strain) GP consists of 629 amino acids and predicts a molecular mass of 69.3 kDa. Partially cleaved by a furin-like convertase to GP1 and GP2 with a predicted molecular mass of 50.9 kDa and 18.4 kDa.
Formulation
Lyophilized from sterile 20 mM Tris, 500 mM NaCl, 10% glycerol, pH 7.5. Please contact us for any concerns or special requirements. Typically 5% – 8% trehalose, mannitol, and 0.01% Tween80 are added as protectants prior to lyophilization. See the specific buffer information on the CoA printout.
Shipping
- In general, recombinant proteins are provided as a lyophilized powder that is shipped at room temperature.
- Recombinant protein bulk packets are provided as frozen liquid.
- They are shipped with blue ice unless otherwise requested by customers.
Stability and storage
Samples are stable for up to twelve months from the date of receipt between -20 ℃ and -80 ℃. Store under sterile conditions between -20-and -80 ℃. It is recommended to take aliquots of the protein for optimal storage. Avoid repeated freeze-thaw cycles.
Reconstitution
A printed copy of the COA with reconstitution instructions is shipped with the products. Please refer to it for detailed information.
General information about the Ebola virus glycoprotein
The fourth gene of the EBOV genome encodes a 16 kDa envelope-bound glycoprotein (GP) and an 11 kDa secreted glycoprotein (sGP). Both GP and sGP have an identical N-terminus of 295 residues, however, they have different C-terminal sequences. Recently, great attention has been paid to GP for vaccine design and isolation of entry inhibitors. GP is a class I fusion protein that assembles as trimers on the viral surface and plays an important role in virus entry and binding.
Mature GP is a disulfide-linked heterodimer made up of two subunits, GP1 and GP2, which are generated from the proteolytic process of the GP precursor (pre-GP) by cellular furin during virus assembly. The GP1 subunit contains a mucin domain and a receptor-binding domain (RBD); the GP2 subunit has a fusion peptide, a helical heptane repeat (HR) region, a transmembrane domain (TM), and a 4-residue cytoplasmic tail. GP1 RBD mediates the interaction of EBOV with the cellular receptor (for example, DC-SIGN / LSIGN, TIM-1, hMGL, NPC1, β-integrins, folate receptor-α, and Tyro3 family receptors), of the which TIM1 and NPC1 are essential for EBOV input; the mucin domain that has N- and O-linked glucans enhances viral binding to cellular hMGL and is involved in protecting key neutralizing epitopes, helping the virus evade immune clearance.
There are large conformational changes of GP2 during membrane fusion, which enhance insertion of the fusion loop into the cell membrane and facilitate the release of the viral nucleocapsid nucleus into the cytoplasm.