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Recombinant Ebolavirus EBOV (subtype Zaire,strain Kikwit-95) Envelope Glycoprotein (GP)  pdf  pdf  pdf


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Synonym

GP1,GP,Envelope glycoprotein,GP2

Source

Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) Envelope Glycoprotein (GP) (Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) GP) Ile 33 - Asp 637 (Accession # AAQ55048.1) was produced in human 293 cells (HEK293) at ACROBiosystems.

Molecular Characterization

Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) GP, containing GP1 and GP2, is fused with a polyhistidine tag at the C-terminus, and has a calculated MW of 67 kDa. The predicted N-terminus is Ile 33. DTT-reduced Protein migrates as 21-23 kDa (GP2-delta) and 110-120 kDa (GP1) in SDS-PAGE .

Endotoxin

Less than 1.0 EU per μg of the Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) GP by the LAL method.

Purity

>85% as determined by SDS-PAGE.

Formulation

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally Mannitol or Trehalose are added as protectants before lyophilization.

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Reconstitution

See Certificate of Analysis for reconstitution instructions and specific concentrations.

Storage

Avoid repeated freeze-thaw cycles.

No activity loss was observed after storage at:
In lyophilized state for 1 year (4oC); After reconstitution under sterile conditions for 3 months (-70oC).

 

SDS-PAGE


Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) Envelope Glycoprotein (GP)
The purity of Ebolavirus EBOV(subtype Zaire,strain Kikwit-95) GP was determined by DTT-reduced (+) SDS-PAGE and staining overnight with Coomassie Blue.
 
 

Background

EBOV encodes seven structural proteins: nucleoprotein (NP), polymerase cofactor (VP35), (VP40), GP, transcription activator (VP30), VP24, and RNA polymerase (L). GP protein contains 160-kDa envelope-attached glycoprotein (GP) and a 110 kDa secreted glycoprotein (sGP). GP is a class I fusion protein which assembles as trimers on viral surface and plays an important role in virus entry and attachment. Mature GP is a disulfide-linked heterodimer formed by two subunits, GP1 and GP2, which are generated from the proteolytical process of GP precursor (pre-GP) by cellular furin during virus assembly . GP1 is responsible for binding to the receptor(s) on target cells. Interacts with CD209/DC-SIGN and CLEC4M/DC-SIGNR which act as cofactors for virus entry into the host cell. GP2 acts as a class I viral fusion protein. GP1,2 mediates endothelial cell activation and decreases endothelial barrier function. sGP seems to possess an anti-inflammatory activity as it can reverse the barrier-decreasing effects of TNF alpha.

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References