Limited proteolysis of herpes simplex virus glycoproteins that occurs during their extraction from vero cells.
Herpes simplex virus glycoproteins extracted from infected Vero cells can be smaller in apparent size than the same viral products extracted from infected HEp-2 cells. Here we show that the differences in size result primarily from limited proteolysis, during or after their extraction, of the viral glycoproteins made in Vero cells. In the absence of appropriate protease inhibitors, both mature and immature forms of four different glycoproteins specified by herpes simplex virus type 2 were significantly smaller (based on electrophoretic mobilities in acrylamide gels) when extracted from Vero cells than when extracted from HEp-2 cells. Inclusion of certain protease inhibitors in the extraction buffer, however, permitted isolation of immature forms from Vero cells that were indistinguishable in size from the immature forms extracted from HEp-2 cells. Under these conditions, the mature forms of glycoproteins B and E were also indistinguishable by electrophoretic sizing from those made in HEp-2 cells, whereas the mature forms of glycoproteins D and F were smaller, indicating the possibility of differences between Vero and HEp-2 cells in the post-translational processing of glycoproteins D and F.
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D018139:Simplexvirus / Q000032:analysis* / Q000378:metabolism; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism*
D006023:Glycoproteins; D011480:Protease Inhibitors; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C038534:glycoprotein gF, herpes simplex virus type 2; D010447:Peptide Hydrolases