title: Cleavage of viral precursor proteins in vivo and in vitro.
abstract: The use of protease inhibitors causes the accumulation of very large polypeptides (polyprotein) in tissue culture cells infected with either poliovirus or echovirus 12. The effectiveness of the inhibitor varies, depending on the cell line chosen. In infected monkey kidney cells, polyprotein is not cleaved when a chymotrypsin inhibitor is added, but in infected HeLa cells a trypsin inhibitor is most effective. Therefore, at least a part of the proteolytic activity is supplied by the host cell. Extracted viral polyprotein can be cleaved in vitro by trypsin or chymotrypsin. As estimated by migration in sodium dodecyl sulfate gels and antigenicity, chymotrypsin cleavage of the poliovirus polyprotein yields fragments which are similar to the in vivo product. The polyprotein is not in soluble form but is attached to a fast-sedimenting, membrane-bound structure. Proteolytic activities in cell extracts were assayed using polyprotein as substrate, and infected and uninfected extracts produced qualitatively dissimilar cleavages.
mesh_terms: D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D002478:Cells, Cultured; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D029822:Enterovirus B, Human / Q000254:growth & development / Q000378:metabolism*; D000882:Haplorhini; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007659:Ketones / Q000494:pharmacology; D007668:Kidney; D008251:Macaca; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D017955:Poliovirus / Q000254:growth & development / Q000378:metabolism*; D011233:Precipitin Tests; D014316:Tritium; D014357:Trypsin / Q000494:pharmacology; D014361:Trypsin Inhibitors / Q000494:pharmacology; D014764:Viral Proteins / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D007659:Ketones; D010455:Peptides; D014361:Trypsin Inhibitors; D014764:Viral Proteins; D014316:Tritium; D002918:Chymotrypsin; D014357:Trypsin