Delta-peptide is the carboxy-terminal cleavage fragment of the nonstructural small glycoprotein sGP of Ebola virus.
In the present study we have investigated processing and maturation of the nonstructural small glycoprotein (sGP) of Ebola virus. When sGP expressed from vaccinia virus vectors was analyzed by pulse-chase experiments using SDS-PAGE under reducing conditions, the mature form and two different precursors have been identified. First, the endoplasmic reticulum form sGP(er), full-length sGP with oligomannosidic N-glycans, was detected, sGP(er) was then replaced by the Golgi-specific precursor pre-sGP, full-length sGP containing complex N-glycans. This precursor was finally converted by proteolysis into mature sGP and a smaller cleavage fragment, Delta-peptide. Studies employing site-directed mutagenesis revealed that sGP was cleaved at a multibasic amino acid motif at positions 321 to 324 of the open reading frame. Cleavage was blocked by RVKR-chloromethyl ketone. Uncleaved pre-sGP forms a disulfide-linked homodimer and is secreted into the culture medium in the presence of the inhibitor as efficiently as proteolytically processed sGP. In vitro treatment of pre-sGP by purified recombinant furin resulted in efficient cleavage, confirming the importance of this proprotein convertase for the processing and maturation of sGP. Delta-peptide is also secreted into the culture medium and therefore represents a novel nonstructural expression product of the GP gene of Ebola virus. Both cleavage fragments contain sialic acid, but only Delta-peptide is highly O-glycosylated.
Volchkova
V A
VA
Institut für Virologie, Philipps-Universität Marburg, Marburg, 35011, Germany.
Klenk
H D
HD
Volchkov
V E
VE
D000595:Amino Acid Sequence; D019281:Dimerization; D029043:Ebolavirus / Q000737:chemistry* / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010446:Peptide Fragments / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D012794:Sialic Acids / Q000378:metabolism; D014162:Transfection; D014764:Viral Proteins
D006023:Glycoproteins; D010446:Peptide Fragments; D012794:Sialic Acids; D014764:Viral Proteins; C110613:secreted glycoprotein, Ebola virus