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Structure-function analysis of the soluble glycoprotein, sGP, of Ebola virus.
- Source :
-
Chembiochem : a European journal of chemical biology [Chembiochem] 2006 Oct; Vol. 7 (10), pp. 1605-11. - Publication Year :
- 2006
-
Abstract
- In addition to the transmembrane protein, GP(1,2), the Ebola virus glycoprotein gene encodes the soluble glycoproteins sGP and Delta-peptide. Two more soluble proteins, GP(1) and GP(1,2DeltaTM), are generated from GP(1,2) as a result of disulfide-bond instability and proteolytic cleavage, respectively, and are shed from the surface of infected cells. The sGP glycoprotein is secreted as a disulfide-linked homodimer, but there have been conflicting reports on whether it is arranged in a parallel or antiparallel orientation. Off-line HPLC-MALDI-TOF MS (MS/MS) was used to identify the arrangement of all disulfide bonds and simultaneously determine site-specific information regarding N-glycosylation. Our data prove that sGP is a parallel homodimer that contains C53-C53' and C306-C306' disulfide bonds, and although there are six predicted N-linked carbohydrate sites, only five are consistently glycosylated. The disulfide bond arrangement was confirmed by using cysteine to glycine mutations at amino acid positions 53 and 306. The mutants had a reduced ability to rescue the barrier function of TNF-alpha-treated endothelial cells--a function previously reported for sGP. This indicates that these disulfide bonds are critical for the proposed anti-inflammatory function of sGP.
- Subjects :
- Amino Acid Sequence
Cells, Cultured
Cysteine chemistry
Disulfides chemistry
Endothelial Cells cytology
Endothelial Cells metabolism
Glycosylation
Humans
Molecular Sequence Data
Mutation
Protein Conformation
Structure-Activity Relationship
Viral Envelope Proteins genetics
Ebolavirus chemistry
Viral Envelope Proteins chemistry
Viral Envelope Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1439-4227
- Volume :
- 7
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Chembiochem : a European journal of chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 16977667
- Full Text :
- https://doi.org/10.1002/cbic.200600223