Biosynthesis of the IFN-γ binding protein of ectromelia virus, the causative agent of mousepox

Ectromelia virus (ECTV), the causative agent of mousepox, expresses an extracellular interferon-γ binding protein (IFN-γBP) with homology to the ligand binding domains of the IFN-γ high affinity receptor (IFN-γR1). Unlike the cellular receptor, the IFN-γBP binds IFN-γ from several species. The IFN-γBP is synthesized early after infection, accumulating in the extracellular milieu as dimers composed of two protein species with Mr of 34.6 or 33.0 kDa. Homodimers are covalently linked by an interchain disulphide bond at position 216. The IFN-γBP has complex N-linked oligosaccharides at positions 41 and 149 as determined by site-directed mutagenesis and glycosidase treatment. Glycosylation at position 41 is required for secretion from mammalian cells and may play a role in the activity of the IFN-γBP. Glycosylation at position 149 is not required for secretion, and the lack of glycosylation at this site does not diminish ligand binding as measured by surface plasmon resonance (SPR) and ELISA.