Coronavirus IBV glycopolypeptides: locational studies using proteases and saponin, a membrane permeabilizer.
[35S]methionine-labelled avian infectious bronchitis virus (IBV) (strain 41) and its purified protein components and virions of IBV-Beaudette were incubated with 10 proteases. Several proteases hydrolysed all or some of the membrane glycopolypeptide (M; Mr 30K) and removed about 1.3K of peptide from the amino-(N-)-terminus plus both glycans, as determined by SDS-polyacrylamide gel electrophoresis. N-terminal analysis of [3H]isoleucine-labelled M after hydrolysis by bromelain revealed that the first nine residues had been removed. After the virions had been permeabilised with saponin, a further 2.5K decrease in molecular weight was produced and this was shown to be from the carboxy-(C-)terminus. When considered with the hydropathicity plot analysis of the amino acid sequence of M (Boursnell, M.E.G. et al., 1984, Virus Res. 1, 303-313) these results suggest that as few as 9-20 N-terminal amino acid residues may protrude at the outer membrane surface and that there is a highly protease sensitive sequence of an estimated 20-25 residues at the C-terminus of M exposed in the lumen of the virion. S2 but not S1 was cleaved to a major glycopolypeptide of approximately 71K by several proteases, and to 76K by trypsin. N-terminal sequencing of the 71K glycopolypeptide revealed that it had the same N-terminus as intact S2. After hydrolysis in the presence and absence of saponin it was concluded that S2 is very sensitive to hydrolysis near its carboxy terminus at residues close to the outer membrane surface.
D000595:Amino Acid Sequence; D003332:Coronaviridae / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D001351:Infectious bronchitis virus / Q000032:analysis* / Q000648:ultrastructure; D008854:Microscopy, Electron; D010447:Peptide Hydrolases / Q000378:metabolism; D012503:Saponins; D014759:Viral Envelope Proteins / Q000032:analysis*; D014771:Virion / Q000378:metabolism
D012503:Saponins; D014759:Viral Envelope Proteins; D010447:Peptide Hydrolases