Interprotein disulfide bonding between F and G glycoproteins of human respiratory syncytial virus.
The envelope glycoprotein G, of human respiratory virus was purified by immunoaffinity chromatography using a monoclonal antibody reacting with G glycoprotein. The purified material was analyzed for its protein patterns and by western blot for its reactivity with specific monoclonal antibodies. In addition to the G specific proteins at 90 and 55 kilodalton (kDa) range, high molecular weight species were coeluted with G protein. Three high molecular weight species were noticed: one (140 kDa) reacting with fusion protein (F) monoclonal antibody and two other species (230 and 195 kDa) reacting with both fusion protein and G protein monoclonal antibodies. The protein reacting only with F monoclonal antibody consists of fusion protein dimer. Western blot and two dimensional gel electrophoretic analysis revealed that each of the other two complexes is composed of two moles of F protein and one mole of G protein. These two complexes differ in their molecular sizes depending on whether G is in the form of 90 or 55 kDa. Upon heat denaturation, fusion protein monomer (70 kDa) is released from the complex, leaving the two complexes, consisting of one mole of F protein and one mole of G protein (160 and 125 kDa species respectively). Disulfide-reducing agents are required to break the monomers of F and G complexes. These results provide a direct evidence for the presence of envelope glycoprotein complexes linked by interprotein disulfide bonding. This may have implications on the structural and functional properties of envelope glycoproteins.
Arumugham
R G
RG
Virology Research, Praxis Biologics, Rochester, New York.
Hildreth
S W
SW
Paradiso
P R
PR
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004220:Disulfides / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000378:metabolism; D015707:HN Protein; D006358:Hot Temperature; D015151:Immunoblotting; D010084:Oxidation-Reduction; D012136:Respiratory Syncytial Viruses / Q000378:metabolism*; D014759:Viral Envelope Proteins; D014764:Viral Proteins
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004220:Disulfides; D006023:Glycoproteins; D015707:HN Protein; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G